Retention Research Articles

Energy Balance and Blood Metabolites Status of Local Sheep Based on Indigofera sp and Sproutbean Waste Ration

<p>This research was undertaken to evaluate energy utilization and blood metabolites status of male Indonesian local sheep involving nutrient balances and using urea space techniques for measurement body composition. Factorial randomized completely design was used in this study using 16 growing male local sheep. Factor A was breed which consisted of 8 Garut sheep (av.BW 14.90 kg) and 8 Jonggol sheep (av.BW 13.60 kg). Factor B was different ration which received of diet containing Indigoferasp. and sproutbean waste. The ration offered were pellets containing CP around 16% and energy 15.6 MJ GE/kg. Energy balance and blood metabolites studies were conducted during three-months trial. Intake, digestibility, urinary and metabolism variables were measured based on total collection method. Methane energy was measured by rusitec technique and methane gas production multiplied by calory equivalent value of methane. Energy retention (RE) was measured by urea space technique. Heat production was calculated from ME-RE. Plasma glucose, cholesterol and urea-N were measured by using spectrophotometry. Results showed that there were signiϐicant differences of energy intakes, digestibilities and metabolism among the rations. Animals fed with sproutbean waste ration has higher energy utilization compared to those fed Indigofera sp ration. There were no signiϐicant different for all blood metabolite variables among breed and its interaction between combination treatments. It is concluded that sproutbean waste in local sheep ration has higher energy utilization and daily gain than Indigofera sp.</p>

Smooth, an hnRNP-L Homolog, Might Decrease Mitochondrial Metabolism by Post-Transcriptional Regulation of Isocitrate Dehydrogenase (Idh) and Other Metabolic Genes in the Sub-Acute Phase of Traumatic Brain Injury.

Traumatic brain injury (TBI) can cause persistent pathological alteration of neurons. This may lead to cognitive dysfunction, depression and increased susceptibility to life threatening diseases, such as epilepsy and Alzheimer's disease. To investigate the underlying genetic and molecular basis of TBI, we subjected w1118 Drosophila melanogaster to mild closed head trauma and found that mitochondrial activity is reduced in the brains of these flies 24 h after inflicting trauma. To determine the transcriptomic changes after mild TBI, we collected fly heads 24 h after inflicting trauma, and performed RNA-seq analyses. Classification of alternative splicing changes showed selective retention (RI) of long introns (>81 bps), with a mean size of ~3,000 nucleotides. Some of the genes containing RI showed a significant reduction in transcript abundance and are involved in mitochondrial metabolism such as Isocitrate dehydrogenase (Idh), which makes α-KG, a co-factor needed for both DNA and histone demethylase enzymes. The long introns are enriched in CA-rich motifs known to bind to Smooth (Sm), a heterogeneous nuclear ribonucleoprotein L (hnRNP-L) class of splicing factor, which has been shown to interact with the H3K36 histone methyltransferase, SET2, and to be involved in intron retention in human cells. H3K36me3 is a histone mark that demarcates exons in genes by interacting with the mRNA splicing machinery. Mutating sm (sm4/Df) resulted in loss of both basal and induced levels of RI in many of the same long-intron containing genes. Reducing the levels of Kdm4A, the H3K36me3 histone demethylase, also resulted in loss of basal levels of RI in many of the same long-intron containing genes. Chromatin immunoprecipitation followed by deep sequencing (ChIP-seq) for H3K36me3 revealed increased levels of this histone modification in retained introns post-trauma at CA-rich motifs. Based on these results, we propose a model in which TBI temporarily decreases mitochondrial activity in the brain 24 h after inflicting trauma, which decreases α-KG levels, and increases H3K36me3 levels and intron retention of long introns by decreasing Kdm4A activity. The consequent reduction in mature mRNA levels in metabolism genes, such as Idh, further reduces α-KG levels in a negative feedback loop. We further propose that decreasing metabolism after TBI in such a manner is a protective mechanism that gives the brain time to repair cellular damage induced by TBI.

Soil labile organic carbon fractions and soil organic carbon stocks as affected by long-term organic and mineral fertilization regimes in the North China Plain

To improve C sequestration in soils and mitigate climate change, it is essential to understand how nutrient management strategies impact on soil organic carbon (SOC) stocks and labile fractions. This study was designed to explore changes in soil bulk density (BD), SOC concentrations, SOC stocks and soil labile organic C fractions (mineralizableC (Cmin), microbial biomass C (MBC), dissolved organic C (DOC), particulate organic C (POC), light fraction organic C (LFOC) and permanganate oxidizable C (KMnO4-C)) under 26-year fertilization regimes in a wheat-maize rotation system in the North China Plain. Soil from the following six treatments was analyzed: (1) Control with no amendment addition (CK); (2) Standard rate of mineral fertilizer treatment (SMF) reflecting local farmers' practice; (3) Standard rate of organic manure treatment (SMA) with total N input equal to SMF; (4) Half the standard rate of mineral fertilizer plus half the standard rate of organic manure treatment (1/2 SMF+1/2 SMA); (5) Double standard rate of mineral fertilizer treatment (DMF); (6) Double standard rate of organic manure treatment (DMA). Results showed that all long-term fertilization regimes significantly decreased BD in topsoil compared to CK except for SMF, with treatments that included organic manure resulting in the lowest BDs. Treatments that included organic manure had significantly higher SOC concentrations and stocks than mineral or unfertilized treatments. The organic manure treatments also had higher concentrations of non-labile C but at the same time a higher proportion of labile C than the mineral or unfertilized treatments. This was confirmed by the carbon management index (CMI) which was significantly increased by organic manure addition. Control and mineral fertilized treatments had higher efficiencies of C retention (RE) from added inputs (crop residues only). Differences in Cmin, POC and KMnO4-C were affected by differences in MA-C, however, changes in rhizodeposition-C, stubble-C and root-C significantly affected DOC, MBC and LFOC. This study demonstrates that fertilization strategies that include organic manure can increase the pool of stable C in the surface soil layer, while at the same time increasing concentrations and proportions of labile C. Organic manure use can therefore contribute to improved nutrient cycling services and higher soil quality in the North China Plain.

IMPLICIT AND EXPLICIT VIDEO FEEDBACK ON LANDING MECHANICS: A RANDOMIZED CONTROLLED TRIAL

BackgroundLower extremity(LE) patterns have been altered using implicit(IF) and explicit(EF) feedback motor learning strategies, however, it remains unclear which strategy may be more effective for injury prevention.ObjectiveTo examine the effects...

Alternative splicing acting as a bridge in evolution.

Alternative splicing (AS) regulates diverse cellular and developmental functions through alternative protein structures of different isoforms. Alternative exons dominate AS in vertebrates; however, very little is known about the extent and function of AS in lower eukaryotes. To understand the role of introns in gene evolution, we examined AS from a green algal and five fungal genomes using a novel EST-based gene-modeling algorithm (COMBEST). AS from each genome was classified with COMBEST that maps EST sequences to genomes to build gene models. Various aspects of AS were analyzed through statistical methods. The interplay of intron 3n length, phase, coding property, and intron retention (RI) were examined with Chi-square testing. With 3 to 834 times EST coverage, we identified up to 73% of AS in intron-containing genes and found preponderance of RI among 11 types of AS. The number of exons, expression level, and maximum intron length correlated with number of AS per gene (NAG), and intron-rich genes suppressed AS. Genes with AS were more ancient, and AS was conserved among fungal genomes. Among stopless introns, non-retained introns (NRI) avoided, but major RI preferred 3n length. In contrast, stop-containing introns showed uniform distribution among 3n, 3n+1, and 3n+2 lengths. We found a clue to the intron phase enigma: it was the coding function of introns involved in AS that dictates the intron phase bias. Majority of AS is non-functional, and the extent of AS is suppressed for intron-rich genes. RI through 3n length, stop codon, and phase bias bridges the transition from functionless to functional alternative isoforms.

Dexamethasone Treatment Leads to Enhanced Fear Extinction and Dynamic Fkbp5 Regulation in Amygdala.

Posttraumatic stress disorder (PTSD) is both a prevalent and debilitating trauma-related disorder associated with dysregulated fear learning at the core of many of its signs and symptoms. Improvements in the currently available psychological and pharmacological treatments are needed in order to improve PTSD treatment outcomes and to prevent symptom relapse. In the present study, we used a putative animal model of PTSD that included presentation of immobilization stress (IMO) followed by fear conditioning (FC) a week later. We then investigated the acute effects of GR receptor activation on the extinction (EXT) of conditioned freezing, using dexamethasone administered systemically which is known to result in suppression of the HPA axis. In our previous work, IMO followed by tone-shock-mediated FC was associated with impaired fear EXT. In this study, we administered dexamethasone 4 h before EXT training and then examined EXT retention (RET) 24 h later to determine whether dexamethasone suppression rescued EXT deficits. Dexamethasone treatment produced dose-dependent enhancement of both EXT and RET. Dexamethasone was also associated with reduced amygdala Fkbp5 mRNA expression following EXT and after RET. Moreover, DNA methylation of the Fkbp5 gene occurred in a dose-dependent and time course-dependent manner within the amygdala. Additionally, we found dynamic changes in epigenetic regulation, including Dnmt and Tet gene pathways, as a function of both fear EXT and dexamethasone suppression of the HPA axis. Together, these data suggest that dexamethasone may serve to enhance EXT by altering Fkbp5-mediated glucocorticoid sensitivity via epigenetic regulation of Fkbp5 expression.

Alpine Skiing With total knee ArthroPlasty (ASWAP): metabolism, inflammation, and skeletal muscle fiber characteristics

We investigated the effect of alpine skiing for 12 weeks on skeletal muscle characteristics and biomarkers of glucose homeostasis and cardiovascular risk factors. Twenty-three patients with a total knee arthroplasty (TKA) were studied 2.9 ± 0.9 years (mean ± SD) after the operation. Fourteen patients participated in the intervention group (IG) and nine in the control group (CG). Blood samples and muscle biopsies were obtained before (PRE) and 7.3 ± 0.8 days after (POST) the intervention, and blood samples again after a retention (RET) phase of 8 weeks. With skiing, glucose homeostasis improved in IG (decrease in fasting insulin, increase in muscle glycogen) but not in CG. Fiber type distribution and size, as well as capillary density and number of capillaries around the fibers (CAF), were not different between the operated and the non-operated leg in either group. The relative number of type I fibers increased with skiing in IG with no change in CG. Inflammatory biomarkers, plasma lipids, and mitochondrial proteins and activity did not change. Alpine skiing is metabolically beneficial and can be used as a training modality by elderly people with TKA.

Effects of variability of practice in music: a pilot study on fast goal-directed movements in pianists.

Variability of Practice (VOP) refers to the acquisition of a particular target movement by practicing a range of varying targets rather than by focusing on fixed repetitions of the target only. VOP has been demonstrated to have beneficial effects on transfer to a novel task and on skill consolidation. This study extends the line of research to musical practice. In a task resembling a barrier-knockdown paradigm, 36 music students trained to perform a wide left-hand interval leap on the piano. Performance at the target distance was tested before and after a 30-min standardized training session. The high-variability group (VAR) practiced four different intervals including the target. Another group (FIX) practiced the target interval only. A third group (SPA) performed spaced practice on the target only, interweaving with periods of not playing. Transfer was tested by introducing an interval novel to either group. After a 24-h period with no further exposure to the instrument, performance was retested. All groups performed at comparable error levels before training, after training, and after the retention (RET) interval. At transfer, however, the FIX group, unlike the other groups, committed significantly more errors than in the target task. After the RET period, the effect was washed out for the FIX group but then was present for VAR. Thus, the results provide only partial support for the VOP hypothesis for the given setting. Additional exploratory observations suggest tentative benefits of VOP regarding execution speed, loudness, and performance confidence. We derive specific hypotheses and specific recommendations regarding sample selection and intervention duration for future investigations. Furthermore, the proposed leap task measurement is shown to be (a) robust enough to serve as a standard framework for studies in the music domain, yet (b) versatile enough to allow for a wide range of designs not previously investigated for music on a standardized basis.

DISPLACEABLE TISSUE: A CLINICAL CHALLENGE FOR A PROSTHODONTIST

A fibrous or flabby ridge is a superficial area of mobile soft tissue affecting the maxillary or mandi bular alveolar ridges. Restoration of such ridges often requires s pecial consideration in impression making. Masticat ory forces can displace this mobile denture-bearing tissue, leadin g to altered denture positioning and loss of periph eral seal. Forces exerted during the act of impression making can res ult in distortion of the mobile tissue. Unless mana ged appropriately by special impression techniques, suc h ‘flabby ridges’ adversely affect the support, ret ention and stability of complete dentures. This paper describe s a technique especially applicable for mandibular edentulous flabby ridges. The choice of the impression materia l, as well as the design of the impression tray, fo cuses on preventing distortion of the displaceable residual ridges during impression making. The use of this te chnique helps in maintaining the contour and capturing the detail of the tissues, as well as in accurately determini ng the extent of the muccobuccal denture extensions.

Energy requirement of growing pigs under commercial housing conditions

Scientifically derived recommendations for the energy supply to growing pigs are generally based on estimates of the metabolisable energy (ME) requirements for maintenance (MEm) and protein (MEp) and fat (MEf) retention. It is supposed that animals are kept within the zone of thermoneutrality and that their physical activity is not elevated. These assumptions might not always be true for practical housing conditions, and it is difficult to quantify the additional energy needed for thermoregulation and physical activity. Hence, at a given ME intake, differences can occur between the actual growth rates and those predicted from the recommendations. To quantify such differences, three trials were carried out under commercial farming conditions with pigs growing from 25 to 120 kg body weight (BW). In each trial, 624 castrated male and female pigs were allocated to four feeding groups distributed over 24 double pens. The rations were provided according to the animals’ feed intake capacity and BW was recorded every three weeks. Protein, fat and energy retention (RE) was derived from carcass composition and BW gain. The difference between ME intake and MEm plus ME required for growth (MEg = RE/kpf) was calculated and seen as the ME required for purposes other than maintenance and growth (MEx). MEx accounted for 2.0%, 17.0% and 21.4% of the animals’ ME intake in Trials 1, 2 and 3, respectively, and was higher in female than in castrated male pigs when related to metabolic BW. It was concluded that total ME requirements of pigs kept under commercial housing conditions can be considerably higher than ME requirements predicted from feeding standards since they usually ignore MEx. MEx can be used as an indicator for the quality of housing systems. Further studies are needed to identify the key factors responsible for MEx to allow for more precise recommendations for the energy supply to commercially raised pigs.

STUDY OF CHEMICAL COMPOSITION OF GARLIC OIL AND COMPARATIVE ANALYSIS OF CO TRIMOXAZOLE IN RESPONSE TO IN VITRO ANTIBACTERIAL ACTIVITY

During the last 20 years, traditional system of medicine has become a topic of global importance. Allium sativum also known as garlic, belong to the Genus Allium is one of the representa tives of this system of medicine. In this study, the garlic oil was extracted by solvent extraction using n - hexane as solvent . The reported percentage yield of oil was 1.1 %, and the specific gravity was 0.90 g/ml. G as C hromatography - M ass S pectrometry anal ysis of oil showed the presence of f our compounds Diallylmonosulfide, Trisulfide methyl 2 propenyl, Diallyl trisulfide, Dimethyl tetrasulfide and their GC retent ion times were 6.2 min utes , 9.0 min utes , 5.9 min utes and 8.1 min utes respectively. The chemical equivalence of drugs plays an important role in their quality assurance. Five different brands of co - trimoxazole were studied for qualitative and quantitative analysis. The result of study showed that samples A - E had 101.29 %, 100.38 %, 99.35 %, 98.65 % a nd 97.31 % of sulfamethoxazole and 99.40 %, 98.69 %, 96.46 %, 89.25 % and 88.67 % of trimethoprim content . Only th ree brands A - C confirmed to official monograph for trimethoprim but two samples D and E showed lower values than standard values. This brin g s the need for drug agencies to monitor the quality of the drugs to check adulteration . The comparison of antibacterial activity of garlic extract and the drug was carried out in - vitro against E - coli , and the result of the study showed that garlic has good a nti - bacterial activity with respect to co - trimoxazole.

Using Pinus uncinata to monitor tropospheric ozone in the Pyrenees

Field metrics were investigated using the conifer species Pinus uncinata for the biomonitoring of tropospheric ozone in the Pyrenees of Catalonia, Spain. The Ozone Injury Index (OII) was investigated piecewise for improvement as a biomonitoring field metric for using sensitive conifer species to monitor tropospheric ozone across variable environmental conditions. The OII employs a weighted average of visual chlorotic mottling (VI), needle whorl retention (RET), needle length (LGT), and crown death (CD). Of note, VI includes subcomponents VI-Amount (% of symptomatic needles) and VI-Severity (% of chlorotic mottling on symptomatic needles) and RET includes the FWHORL subcomponent (average fraction of needles retained per whorl). All components and subcomponents of the OII correlated better to multiple year ozone exposure compared to single year ozone exposure measurements. VI-Severity and FWHORL modeled over half the variability of the three year average of ambient ozone concentrations (P<0.0001, R2=0.53, RMSE=2.73). Combining the biomonitoring metrics with GIS models related to landscape-scale variability in plant water relations resulted in considerable improvement in the ozone exposure model explanatory power (P<0.0001, R2=0.90, RMSE=1.35) including the parameters VI-Amount, VI-Severity, elevation, slope and topographic curvature.

DEVELOP A NEW RP-HPLC METHOD AND VALIDATION FOR QUANTITATIVE DRUG CONTENT DETERMINATION OF EZETIMIBE FROM MARKETED FORMULATED TABLETS

This study has done to develop a cheap, reliable and stable method for the drug content determination of Ezetimibe from its formulated tablets. A binary reversed – phase (C 18 - Column) high performance liqui d chromatograph was used for the analysis. An isocratic method was applied with the mobile phase acetonitrile and water in the ratio of 75:25 (by volume). For detection the Ezetimibe analyte UV - Detector was used at the wave length 230 nm. The average reten tion time of Ezetimibe was 3.30 min utes . The flow rate was 1.0 mL / min. The method was linear with regression coefficient (R 2 ) 0.994. The limit of detection (LOD) and limit of quantification (LOQ ) were calculated 0.04 and 0.12 µg / mL. Average inter - day a nd intra - day recovery percentage were computed 98.44 % and 99.62 % with coefficient variation percentage 3.07 and 0.75 %. Recovery results were showed good accuracy of the method. Hence, this method validation was statistically significant.

Matrix structure selection in the microparticles of essential oil oregano produced by spray dryer

The goal of this work was to select the best combination of encapsulants for the microencapsulation of oregano essential oil by spray dryer with the addition of Arabic gum (AG), modified starch (MS) and maltodextrin (MA). The simplex-centroid method was used to obtain an optimal objective function with three variables. Analytical methods for carvacrol quantification, water activity, moisture content, wettability, solubility, encapsulation efficiency (ME) and oil retention (RT) were used to evaluate the best combination of encapsulants. The use of AG as a single wall material increased ME up to 93%. Carvacrol is the major phenolic compound existent in the oregano essential oil. Carvacrol exhibits a maximum concentration of 57.8% in the microparticle with the use of 62.5% AG and 37.5% MA. A greater RT (77.39%) was obtained when 74.5% AG; MS 12.7% and 12.7% MA were applied, and ME (93%) was improved with 100% of gum.

On the interrelation between the distribution constant and the retention factor in liquid chromatography

The relationships that associate the equilibrium distribution constant and retention factor in high� performance liquid chromatography are analyzed critically. The K c = kV M /v a + 1 relationship suggested by Yu.A. El'tekov is derived without using the relationship between the Henry adsorption constant and the reten� tion factor. The incorrectness of using another relationship, Kc = k/ϕ, to calculate the distribution constant, which is determined by ignoring the excess character of the retention factor measured experimentally when deriving it, is demonstrated. It is proposed that the retention factor be determined by the ratio of the excess amount of adsorbate in an adsorption layer to the amount of adsorbate in bulk solution.

How common is colonic elongation in children with slow-transit constipation or anorectal retention?

PurposeColonic elongation is reported as a possible cause for slow colonic transit, as it is observed in patients with slow-transit constipation (STC). This study aimed to determine the frequency of colonic elongation in children with STC or anorectal retention using radioimaging. We hypothesized that transverse colon elongation may occur in patients with STC, whereas sigmoid colon elongates in patients with anorectal retention. MethodsNuclear transit scintigraphy performed for chronic constipation (1999-2011) was analyzed qualitatively for elongated transverse colon or sigmoid colon. Three major colonic transit patterns were identified: slow transit in the proximal colon (STC), normal proximal colonic transit with anorectal retention (NT-AR), and rapid proximal transit ± anorectal retention (RT). χ2 Test was used for statistical analysis (P < .05 significant). ResultsFrom 1999 to 2011, 626 children had nuclear transit scintigraphy. Transverse colon elongation occurred more frequently in STC (73/322, or 23%) compared with NT-AR (9/127, or 7%) and RT (5/177, or 3%; P < .0001). Sigmoid colon elongation was equally common in NT-AR (8/127, or 6%) compared with RT (10/177, or 6%) and STC (14/322, or 4%; P < .9). ConclusionTransverse colon elongation is more common in STC (23%), whereas sigmoid colon elongation is not more common in anorectal retention. Colonic elongation may be the cause or the result of the underlying slow colonic transit.

A myocardial ischemia- and reperfusion-induced injury is mediated by reactive oxygen species released from blood platelets

In recent experimental studies, blood platelets have been found to exhibit some cardiodepressive effects in ischemic and reperfused guinea pig hearts independent of thrombus formation. These effects seemed to be mediated by reactive oxygen species (ROS). However, the source of these ROS – platelets or heart – remained still unknown. Isolated, buffer-perfused and pressure-volume work performing guinea pig hearts were exposed to a low-flow ischemia (1 ml/min) of 30 min duration and reperfused at a constant flow of 5 ml/min. Human thrombocytes were administered as 1 min bolus (20 000 thrombocytes/µl perfusion buffer) in the 15th min of ischemia or in the 1st or 5th min of reperfusion in the presence of thrombin (0.3 U/ml perfusion buffer). Recovery of external heart work (REHW) was expressed as ratio between postischemic and preischemic EHW in percent. Intracoronary platelet retention (RET) was quantified as percent of platelets applied. In a second set of experiments, thrombocytes were incubated with 10 µM of the irreversible NADPH oxidase blocker diphenyliodonium chloride and washed twice, thereafter, and administered according to the same protocol as described above. Hearts exposed to ischemia and reperfusion in the presence of thrombin but without application of platelets served as controls. Controls without application of platelets did not reveal a severe compromisation of myocardial function (REHW 85.5 ± 1%). However, addition of platelets during ischemia or in the 1st or 5th min of reperfusion led to a significant reduction of REHW as compared with controls (REHW 62.4 ± 6, 53.9 ± 3, 40.5 ± 3, respectively). Application of platelets pretreated with diphenyliodonium chloride did not reveal any cardiodepressive effects being significantly different from controls without platelet application. Moreover, treatment of platelets with diphenyliodonium chloride did not significantly decrease intracoronary platelet retention. In conclusion, these results demonstrate that cardiodepressive effects of human thrombocytes in ischemic and reperfused guinea pig hearts are mediated by ROS released from thrombocytes and not the heart.

A comparative electrophysiological study of regulatory components of working memory in adults and seven- to eight-year-old children: An analysis of coherence of EEG rhythms

Coherence at the frequency of θ, α, and β EEG rhythms was analyzed in 14 adults and 23 children of 7−8 years old while they performed cognitive tasks requiring an involvement of working memory (WM). We used the pair matching paradigm in which subjects had to match a pair of stimuli shown in succession in the central visual field. The pairs of verbal and visuospatial stimuli were mixed together and presented in a pseudo random order. Each pair was preceded by a warning signal that did not specify a modality of upcoming stimuli. We analyzed EEG segments recorded (i) in the rest condition, (ii) prior to the first (reference) stim� ulus (maintenance of nonspecific voluntary attention), and (iii) prior to the second (test) stimulus (retention of information in WM). In the present study we focused on the regulatory functional components of WM, and therefore, the stimulus modality has not been taken into account. In adults, maintaining nonspecific vol� untary attention was accompanied by an increase of the strength of θ�related functional coupling between medial areas of the frontal cortex and temporal cortical zones and by a strengthening of local β�related func� tional connectivity in the frontocentral areas of the cortex. In children, no such increase was found for θ rhythm; for β rhythm the increase was limited to several shortrange functional links. In adults, the reten� tion of information in WM was accompanied by the growth in α coherence in distant frontoparietal links, predominantly in the right hemisphere, while in children information retention was accompanied by the growth of θ coherence in the inferiotemporal and parietal co rtical regions. The results of the study point to a relative immaturity of the mechanisms of executive control of WM in children of 7-8 years old.

Biodistribution and pharmacokinetics of variously sized molecular radiolabelled polyethyleneiminomethyl phosphonic acid as a selective bone seeker for therapy in the normal primate model.

An ideal radiopharmaceutical for the treatment of neoplastic and inflammatory (benign) bone disease would be a radiolabelled compound that predominantly accumulates in bone lesions with limited access to normal bone and other organs. Neoplastic tissue's abnormal blood supply (increased permeability) and lack of lymphatics will selectively accumulate radiolabelled macromolecules. This enhanced permeability and retention effect forms the basis of this study, using various molecular sizes of the radiolabelled macromolecule polyethyleneiminomethyl phosphonic acid (PEI-MP) for increased selectivity of the bone-seeking radiopharmaceutical. PEI-MP was synthesized by condensation of polyethyleneimine, phosphonic acid and formaldehyde, followed by fractionation into different molecular sizes by membrane ultrafiltration. Labelling efficiency to 99mTc (as radiotracer) was approximately 99% with complexes stable for 24 h. The pharmacokinetics and biodistribution of various 99mTc-PEI-MP fractions were investigated using 4 experimental baboons (Papio ursinus) per fraction. Scintigraphy was performed on the baboons under general anaesthesia of pentobarbital i.v. After an i.v. bolus of 99mTc-PEI-MP (approximately 185 MBq) both dynamic studies (30 x 1 min frames), and static studies (2 min acquisition every hour for 4 h) were done, as well as blood samples and urine collected. From the results macromolecules with sizes ranging between 30-300 kDa were characterized by excessive liver (21%-57% retained activity) and kidney (40% retained activity) uptake and accompanying long residing times (t1/2 up to 24 h). The percentage bone uptake averaged at 8% for these particles excluding sizes 100-300 kDa where very little bone uptake was seen (< 1%). In this case the blood clearance was also slow (t1/2 approximately 2 h). The fraction size 10-30 kDa had comparatively low accumulation and short residence times in the liver and kidneys (resp. 20%, t1/2 = 22 +/- 4 min; 17.5%, t1/2 = 20 +/- 3 min) and although the bone uptake of 18% in this case was high, it is still low for a bone-seeking agent. These particles cleared the blood with t1/2 = 25 +/- 2 min and seemed suitable for labelling with a therapeutic radioisotopic agent.

Cyto-nuclear discordance in the genetic relationships among Tunisian fig cultivars ( Ficus carica L.): Evidence from non coding trnL– trnF and ITS regions of chloroplast and ribosomal DNAs

Genetic diversity studies were conducted to evaluate intra-specific relationships in Tunisian fig (Ficus: Moraceae) using sequences of the ITS regions of nuclear ribosomal DNA and the chloroplast non-coding region trnL– trnF ( trnL intron, and trnL [UAA]–exon– trnF [GAA] intergenic spacer). All data sets suggest that the sequences obtained showed variations either on their lengths or on their nucleotide compositions. The mean size of these nuclear and cytoplasmic non-coding regions is 697.5 and 1035 base pairs for ITS and trnL– trnF chloroplast DNA, respectively. Our results suggest that the substitution rate estimated for the ITS sequences of nuclear DNA is greater than the one unregistered for the non-coding regions of chloroplast DNA. Therefore, chloroplast DNA shows more homoplasy than the nuclear ribosomal DNA. In fact, the consistency (CI) and retention (RI) indexes calculated for nuclear and cytoplasmic DNAs show values of 0.420 and 0.490 for chloroplast DNA and 0.573 and 0.387 for ribosomal DNA, respectively. The nuclear DNA has more resolution of genetic relationships between cultivars. In addition, the result suggests that the cultivars studied are clustered independently from their geographical origin and the male trees did not thoroughly diverge from the common figs. The discordance between chloroplast and the nuclear topology is revealed. The lack of congruence between the two data sets may be a result of hybridization or introgression. Our result showed that the nuclear and cytoplasmic sequences are useful for germplasm discrimination as well as for investigation of patterns of variation in fig.