A method for monitoring intensity during aquatic resistance exercises

The aims of this study were (i) to check whether monitoring of both the rhythm of execution and the perceived effort is a valid tool for reproducing the same intensity of effort in different sets of the same aquatic resistance exercise (ARE) and (ii) to assess whether this method allows the ARE to be put at the same intensity level as its equivalent carried out on dry land. Four healthy trained young men performed horizontal shoulder abduction and adduction (HSAb/Ad) movements in water and on dry land. Muscle activation was recorded using surface electromyography of 1 stabilizer and several agonist muscles. Before the final tests, the ARE movement cadence was established individually following a rhythmic digitalized sequence of beats to define the alternate HSAb/Ad movements. This cadence allowed the subject to perform 15 repetitions at a perceived exertion of 9-10 using Hydro-Tone Bells. After that, each subject performed 2 nonconsecutive ARE sets. The dry land exercises (1 set of HSAb and 1 set of HSAd) were performed using a dual adjustable pulley cable motion machine, with the previous selection of weights that allowed the same movement cadence to be maintained and the completion of the same repetitions in each of the sets as with the ARE. The average normalized data were compared for the exercises in order to determine possible differences in muscle activity. The results show the validity of this method for reproducing the intensity of effort in different sets of the same ARE, but is not valid for matching the same intensity level as kinematically similar land-based exercises.     

5-Hydroxytryptophan: A method for monitoring human plasma content during clinical administration

A method is described for the assay of 5-hydroxytryptophan (5-HTP) in human blood plasma following administration of the compound. The procedure involves extracting the amino acid into butanol, returning the 5-HTP to an aqueous phase, and separating it from other interfering 5-hydroxyindoles on a column of Dowex 50W-X4, a strong cation-exchange resin. The concentration of 5-HTP in the buffer eluate is determined spectrofluorometrically in 3 N HCl; 25 ng/ml plasma can be detected. Using this method, it was possible to determine the time-course of plasma 5-HTP concentrations during the 24-h period after a single 300-mg dose. 5-HTP is being utilized in clinical trials for several psychiatric and neurological disorders.     

A novel method for monitoring monoclonal antibody production during cell culture

We describe a new format for surface-based fluoroimmunoassays that allows detection of biomolecule interactions without separation steps. The bioactive layer was immobilized on the surface of a glass substrate covered with silver islands that provide optical amplification of the distinctive fluorescence signal from bound probes when compared to unbound probes. The technique used was phase-modulation fluorometry that allows sensitive detection of bound probes with a very short lifetime in the presence of excess free probes in solution. The new method was applied to assay monoclonal antibody production during cell culture. Excellent agreement was found between the new method and ELISA analysis of hybridoma cell culture samples. It is predicted that the near real time monitoring of protein products during bioprocessing will be possible with the described technology.     

A method for continuous monitoring of meconium in the amniotic fluid during labour

In about 10% of pregnancies overall, the fetus discharges meconium (its bowel contents) into the amniotic fluid during labour. In about 10% of cases where meconium is passed, the fetus gasps, inhaling the sticky meconium into the upper respiratory tract. After birth, the meconium blocks the air passages in the lungs, impairing gas exchange — meconium aspiration syndrome (MAS). Up to 20% of infants suffering from MAS die and recently published studies have shown a long-term effect of MAS in causing cough and wheeze. The risk of meconium aspiration is thought to be increased by intrauterine hypoxia. At present, meconium is only noticed at birth or occasionally when amniotic fluid leaks past the presenting part of the fetus. A method has been developed which measures absolute meconium concentration with a 99% prediction interval of ± 30 gl⁻¹; allows monitoring of the rate of appearance of meconium linearly with a nonlinearity of 5%, and differentiates between meconium and blood. The method uses the ratio of the intensity of back-scattered light from the amniotic fluid at 700 and 415 nm, the latter being near the peak of light absorption by meconium and the former a reference value. The ratio is also affected by the presence of blood. However, blood has specific absorption peaks at 540 and 575 nm from which it can be detected (the presence of blood is also a significant abnormality, and is relatively uncommon). The measurement method could easily be integrated into an optical sensor mounted onto an intrauterine probe. The measurement of back-scattered light at 415, 540 and 700 nm would allow continuous monitoring of meconium which could provide valuable information for the study of the pathophysiology of meconium passage in utero during labour and allow preventative measures to be developed.     

A method for maintaining normoglycemia during labour and delivery in insulin-dependent diabetic women.

The effectiveness of combining the subcutaneous administration of short- and intermediate-acting insulin with the intravenous infusion of glucose in maintaining normoglycemia during labour and delivery in insulin-dependent diabetic women was tested. Fifty women were given intermediate-acting insulin twice daily in doses that were fractions of their usual dose, based on the projected duration of labour. In addition, they were given regular (i.e., short-acting) insulin every 6 hours, the dose being 1% of their total daily insulin dose for every increase of 10 mg/dl above 100 mg/dl (5.6 mmol/l) in the plasma glucose level 1 hour previously; the levels were measured every 3 hours. All the patients were fasting and received a basal intravenous infusion of 6 g/h of glucose; the rate of infusion was increased by 1 g/h for every decrease of 10 mg/dl in the plasma glucose level below 100 mg/dl. The mean plasma glucose levels (+/- standard deviation) were 90 +/- 46 mg/dl after 3 hours of labour, 92 +/- 35 mg/dl after 6 hours, 97 +/- 49 mg/dl after 9 hours and 107 +/- 65 mg/dl after 12 hours. With only one exception, in a premature infant, the 5-minute Apgar scores were identical to those of the infants of nondiabetic women.     

A simple method for monitoring chromatography column eluates for laccase activity during enzyme purification

A simple and economical method is described that allows rapid detection of laccase activity in chromatography column fractions during enzyme purification. Aliquots of column eluants are applied to filter paper coated with 2,2-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) containing a numbered grid, and incubated at ambient temperature for 20 min. Indications of enzyme activity are simply observed by a colour change. This method avoids having to manually assay each fraction of a chromatographic run for enzyme activity.     

A method for continuous monitoring of phosphorylase b activity during glycogen degradation and synthesis

Phosphorylase b (1,4-α-glucaniorthophosphate glucosyltransferase, EC. 2.4.1.1) catalyzes the glycogen synthesis and degradation reaction. It is the latter which is the physiological function of the enzyme. Its action may be described by a random order bi-bi kinetic mechanism. The rate-limiting step is an interconversion between ternary complexes (1).Activity determination can be carried out in the direction of glycogen synthesis by measuring the amount of inorganic phosphate (P_j) released during the reaction (2). This method requires termination of the enzyme action after an appropriate time interval and only yields information about the total amount of the product without giving details of the time course of the process. In the reverse direction, the glycogen degradation can be followed in a more complicated and indirect way by coupled reactions of additional enzymes (3).The present communication describes a convenient method for continuous monitoring of the reaction in both directions with a fluorescence spectrometer.     

A novel method for monitoring protoplast fusion

Summary Two fluorescent compounds, scopoletin and carboxyfluorescein, have been used to label both tissue culture and leaf mesophyll cells and protoplasts. The compounds localized within the vacuoles of cells in approximately 15 hours. They remained in the vacuole during cell wall digestion, and fluorescence was observable for several hours after protoplast release. A one day pulse of these fluorescent labels had no deleterious effect on the growth of cells or protoplasts. When morphologically indistinguishable protoplasts were labeled and treated with polyethylene glycol, multicolored fluorescent fusion products were observable. These fluorescent labels provide a convenient method for selection of heterokaryon fusion products of whole plant and tissue culture cell protoplasts.     

A survey on management and housing of peri-parturient dairy cows and their calves

Housing and management around the time of calving impact dairy cow behaviour, health and welfare, but little is known about current practice. The aim was to provide an overview of current calving practice and the study describes the main calving housing and management based on replies to an online questionnaire by 42 dairy cattle experts in 28 countries, or regions, in Europe, Canada and USA. The survey suggests that in the majority of countries and regions included in this study, dairy cows typically calve in indoor calving facilities; either individual pens, group pens or a system where the cow is moved from a group pen into an adjacent individual pen before calving. Regarding individual calving pens, the survey suggests that in the majority of countries and regions included in this study, most pens have open sides and offer cows no opportunity to isolate, although research shows that a secluded corner of an individual pen creates a preferred calving site. Further, the survey suggests that when cows calve in individual calving pens or tie-stalls, they are often moved there with signs of imminent calving, although research shows that this practice increases the duration of calving and it is recommended to move cows before their expected calving time. Regarding group pens, none of the 42 respondents replied that group pens typically offer cows the opportunity to isolate at calving. Recent research suggests that when cows calve in a secluded area of a group calving pen, this reduces the risk of failure of passive transfer of immunity. Regarding calving facilities where group pens are combined with adjacent individual pens, this was reported to be the most typical in 10 of the 24 countries and regions with indoor calving sections covered by the survey. The same concerns regarding when the cow is moved from the group pen to an individual pen apply, as outlined above. Irrespective of pen type, the most frequently reported surface was deep bedded straw and the most frequent type of separation between pens was open sides. Cow-calf separation within 12 hours of birth, and thereafter individual housing of calves combined with milk feeding via a teat bucket or bar was indicated the most frequent management. The survey presents experts’ evaluations of current practice of housing and management of peri-parturient dairy cows and their calves, and suggests that there is a discrepancy between current calving management and housing and recommendations based on recent research.     

Effect of rumen-protected methionine supplementation to beef cows during the periconception period on performance of cows,calves, and subsequent offspring

Maternal nutrition affects the development of the fetus and postnatal performance of the calf. Methionine may play a critical role in developmental programming and is likely deficient in beef cows fed low-quality forage. The objective of this study was to determine the effect of metabolizable methionine supply to lactating beef cows during the periconception period on performance of cows, calves, and subsequent offspring. This project involved two consecutive production cycles commencing at calving in which dietary treatments were fed to cows during the periconception period along with measurements on cows and initial calves in Production Cycle 1, and measurements on subsequent calves in Production Cycle 2. Brangus-Angus crossbred lactating beef cows (N = 108; age = 6.4 (2.8) year) were stratified by previous calving date and assigned to one of three supplements: (1) control, molasses plus urea at 2.72 kg/day as fed, (2) fishmeal, 2.27 kg/day molasses plus urea plus 0.33 kg/day as fed of fishmeal, and (3) methionine, 2.72 kg/day of molasses plus urea plus 9.5 g/day of 2-hydroxy-4-(methylthio)-butanoic acid. Cows were fed supplements and low-quality limpograss (Hemarthria altissima) hay while grazing dormant bahiagrass (Paspalum notatum Flüggé) pastures during the 115-day periconception period from December 2014 to April 2015 in Production Cycle 1 only. Body weight change and milk yield of cows were measured during the periconception period in Production Cycle 1. Body weight of calves was measured at birth and weaning in both production cycles. Following weaning in Production Cycle 2, eight subsequent steer calves per treatment were individually housed for a 42-day metabolism experiment. Treatment did not affect (P > 0.10) BW change of cows, but cows fed methionine tended (P = 0.09) to produce more energy-corrected milk than control and fishmeal. Treatment did not affect (P > 0.10) 205-day adjusted weaning weight of calves in either production cycle. During the metabolism experiment, subsequent calves from dams fed fishmeal and methionine gained faster (P < 0.05) and had greater (P < 0.05) gain:feed than control calves. Methionine calves tended (P = 0.06) to have greater apparent total tract NDF and ADF digestibility and lesser (P < 0.05) blood glucose concentration than control and fishmeal calves. These data indicate that maternal methionine supply during the periconception period plays an important role in programming future performance of the offspring.     

A robust in vivo positive-readout system for monitoring siRNA delivery to xenograft tumors

Delivering small interfering RNA (siRNA) to tumors is the major technical hurdle that prevents the advancement of siRNA-based cancer therapy. One of the difficulties associated with the development of clinically relevant delivery systems is the lack of reliable tools for monitoring siRNA delivery to tumors in vivo. We describe here a novel, positive-readout system where siRNA-mediated target knockdown elicits a rapid and robust increase of reporter activity. Using the positive-readout system, we created (1) β-galactosidase-based tumor models that allow the detection of target knockdown in 1%-2% of tumor cells and can distinguish between tumor areas where effective target knockdown occurs versus tumor areas that are not accessible to delivery, and (2) luciferase-based tumor models that allow the quantitative assessment of a large number of delivery systems. Using these positive-readout models, we screened a number of literature-described siRNA delivery systems and identified lipid nanoparticles as a promising delivery platform for siRNA-based cancer therapy.     

Birth delivery method affects expression of immune genes in lung and jejunum tissue of neonatal beef calves

Background

Caesarean section is a routine veterinary obstetrical procedure employed to alleviate dystocia in cattle. However, CS, particularly before the onset of labour, is known to negatively affect neonatal respiration and metabolic adaptation in humans, though there is little published information for cattle. The aim of this study was to investigate the effect of elective caesarean section (ECS) or normal trans-vaginal (TV) delivery, on lung and jejunal gene expression profiles of neonatal calves.

Results

Paternal half-sib Angus calves (gestation length 278 + 1.8 d) were delivered either transvaginally (TV; n?=?8) or by elective caesarean section (ECS; n?=?9) and immediately euthanized. Lung and jejunum epithelial tissue was isolated and snap frozen. Total RNA was extracted using Trizol reagent and reverse transcribed to generate cDNA. For lung tissue, primers were designed to target genes involved in immunity, surfactant production, cellular detoxification, membrane transport and mucin production. Primers for jejunum tissue were chosen to target mucin production, immunoglobulin uptake, cortisol reaction and membrane trafficking. Quantitative real-time PCR reactions were performed and data were statistically analysed using mixed models ANOVA. In lung tissue the expression of five genes were affected (p?<?0.05) by delivery method. Four of these genes were present at lower (LAP, CYP1A1, SCN11α and SCN11β) and one (MUC5AC) at higher abundance in ECS compared with TV calves. In jejunal tissue, expression of TNFα, Il-1β and 1 l-6 was higher in ECS compared with TV calves.

Conclusions

This novel study shows that ECS delivery affects the expression of key genes involved in the efficiency of the pulmonary liquid to air transition at birth, and may lead to an increased inflammatory response in jejunal tissue, which could compromise colostral immunoglobulin absorption. These findings are important to our understanding of the viability and management of neonatal calves born through ECS.

     

A novel method for activity monitoring in small non-human primates

Patterns of spontaneous activity are valuable reflections of well-being in animals and humans and, because of this, investigations have frequently incorporated some form of activity monitoring into their studies. It is widely believed that activity monitoring, alongside assessments of general behaviour, should be included in initial CNS safety pharmacology screening. As the number of marmoset studies having actimetry as their focus, or as an adjunct, is increasing, we wished to evaluate an alternative approach to those commonly used. The method is based on miniaturized accelerometer technologies, currently used for human activity monitoring.Actiwatch-Minis were used to monitor the activity of two groups of differently housed marmosets for 14 consecutive days. Group A consisted of four mixed-sex pairs of animals and group B comprised eight group-housed males. Activity profiles were generated for weekday and weekend periods. The devices captured quantifiable data which showed differences in total activity between the two differently housed groups and revealed intragroup variations in the temporal spread of activity between weekdays and weekends. The Actiwatch-Mini has been shown to generate retrospective, data-logged activity counts recorded from multiple animals in a single arena by means of non-invasive monitoring.     

Genotype by environment interaction for livability of dairy calves from first parity cows

Death of calves around parturition is a matter of concern for dairy farmers. Relatively high stillbirth rates and unfavourable trends have been reported for Holstein heifers in the Netherlands and several other countries. In our study, we investigated herd differences, genetic parameters and genotype by environment interaction for heifer calf livability. A large dataset with data from calvings between 1993 and 2012 of Dutch dairy farms was used. There were considerable differences between herds in livability of calves from heifers, with averages ranging from 74% to 95%. Both herds with relatively high and low averages showed the same negative trend between 1993 and 2012, with largest declines in herds with the lowest averages. We found that heritability and genetic variation of first parity livability were substantially larger in herd environments where the likelihood of stillbirth was high v. environments where stillbirth was at a low level. The genetic correlations between herd environment levels were all very close to unity, indicating that ranking of sires was similar for all environments. However, for herds with a relatively high stillbirth incidence selecting sires with favourable breeding values is expected to be twice as profitable as in herds with a relatively low stillbirth incidence.     

Pharmacokinetic monitoring during aminoglycoside treatment: the optimal method for individualizing the dosage of tobramycin

The results of tobramycin concentration monitoring in 33 patients with nonspecific pulmonary infections showed a marked individual variability of the antibiotic blood levels and model-independent pharmacokinetic parameters: total clearance, steady-state volume of distribution and mean residence time whose values were distributed log-normally. Adjusting of the tobramycin dosage by the individual values of the clearance (three-point method, by concentrations 1 h (C1), 3 h (C3) and 6 h (C6), after intramuscular single administration of the antibiotic and one-point method, by C3, after repeated administrations of the antibiotic) provided by the end of a 7-day course a 1.7-fold decrease in the individual ranges of the antibiotic concentration as compared to those without the dosage adjusting. Retrospective analysis revealed that reliable individual dosing of tobramycin was provided with the simplest one-point method when the only blood specimen was collected 3 hours after the injection, i.e. the time interval inversed to the elimination rate constant. According to this method individual doses Dind were calculated by the equation Dind = DpopCpop/Cind, where pop was the population value of D and C. The values of Dind estimated in such a way did not practically differ from those estimated with the more complicated two-point (by C1 and C6) and three-point methods. Application of the equation to the tobramycin "maximum" concentration C1 or the "minimum" one (toward the end of the dosing interval, C6) resulted in less accurate and unbiased estimation of Dind.     

A new method for targeted drug delivery using polymeric microcapsules

Recent research and clinical evidence suggest that thalidomide could potentially be used to treat inflammation associated with Crohn's disease. However, systemic side effects associated with large doses of this drug have limited its widespread use. Treatment, with thalidomide would prove more efficacious if the drug could be delivered directly to target areas in the gut, thereby reducing systemic circulation. Microcapsule encapsulation could enable direct delivery of the drug. To assess the latter, we designed and tested drug-targeting release characteristics of alginate-poly-l-lysine-alginate (APA) microcapsules in simulated gastrointestinal environments. The results show that APA capsules enabled delivery of thalidomide in the middle and distal portions of the small intestine. We also compared the APA membrane formulation with an earlier designed alginate chitosan (AC) membrane thalidomide formulation. The results show that both APA and AC capsules allow for successful delivery of thalidomide in the gut and could prove beneficial in the treatment of Crohn's disease. However, further research is required.     

A kinetic method for measuring functional delivery of amphotericin B by drug delivery systems.

The human toxicity of amphotericin B can be considerably reduced by associating the drug with liposomes of varying lipid compositions. Some lipid compositions are much more effective than others. We show that a simple kinetic fluorescence assay using pyranine as an indirect probe of amphotericin-induced K+ currents may be used to study different liposomal drug delivery systems in vitro. We find that lipid mixtures composed of DMPC/DMPG/amphotericin at a 7:3:1 mole ratio show very slow functional delivery with a preference for ergosterol over cholesterol-containing membrane vesicles. On the other hand, amphotericin delivered from egg phosphatidylcholine liposomes lead to 100-fold increases in K+ leakage at one-fifth the amphotericin concentration of the 7:3:1 system. The egg phosphatidylcholine system as well as micellar amphotericin also show a slight selectivity towards cholesterol-containing vesicles over ergosterol. These results are consistent with previous clinical and in vitro cellular studies and this technique may prove valuable in screening of other delivery systems.