Lying patterns of high producing healthy dairy cows after calving in commercial herds as affected by age,environmental conditions and production

An animal expresses its physiological and well-being status by its behaviour. Changes in behaviour can be associated with health, production or well-being problems and therefore with the profitability of the farm. The objectives of the present study were to analyse lying patterns of healthy cows, collected with a commercial behaviour sensor, in early lactation in relation to environmental conditions, age of the cow and production performance. In future, these results may be used as a ‘baseline’ for detection of alterations in behaviour that indicate health problems. The study involved 210 healthy multiparous Israeli Holstein cows in three commercial dairy farms. Only healthy cows during the first 28 days after calving were included in this study. Data were analysed in relation to calving season, age of cows and correlation between milk production and lying time.The results show that lying time increased significantly with age and is significantly (P < 0.05) higher in winter than in summer (summer lactation 2: 491 ± 17 min/day (mean ± SD), summer lactation 3 and more: 520 ± 25 min/day, winter lactation 2: 531 ± 25 min/day, winter lactation 3 and more: 579 ± 38 min/day).The proportion of positively and negatively milk production and lying time correlated cows is affected by calving season.This study indicates that behaviour variables in early lactation are affected by calving season, lactation number and type of correlation between milk production and lying time.     

A brief report on intra-species aggressive biting in a goat herd

This study aimed at analysing the effects of age, horn and places on biting and butting as intra-species aggressive activities. Seventy-two Turkish Saanen goats were used as animal material and 22,686 aggressive behaviours were recorded in different places from a total of 118 h direct observation. Of the observations of aggressive behaviours, 32.7% was biting behaviour and the rest was butting behaviour. The frequency of biting behaviour in 3 or more years old goats was significantly higher than that of biting behaviour in 1 and 2 years old animals (P ≤ 0.01). The observations revealed that horned or hornless goats exhibited biting behaviour; however the frequency of biting behaviour in hornless goats was 2.38 times higher than in horned goats (P ≤ 0.01). Biting and butting behaviours were found to be well correlated with the area of places (P ≤ 0.01). As the area of places got narrowed, the frequency of biting increased. However, such a trend was not observed in butting behaviour. The frequency of butting behaviour again increased with the increase in social hierarchy (P ≤ 0.01), whereas the frequency of biting behaviour was not affected by social hierarchy (P = 0.30). In conclusion, intra-species biting behaviour, which is thought to have developed as a result of horn absence, should be questioned whether this is unique to the herd or to the genotype.     

Seed storage and germination in Kumara plicatilis,a tree aloe endemic to mountain fynbos in the Boland,south-western Cape,South Africa

Seed storage under appropriate conditions is a relatively inexpensive means of safeguarding plant genetic material for ex situ conservation. Post-storage germination trials are used to determine the viability of stored seeds, and hence the efficacy of the particular storage treatment. Kumara plicatilis (= Aloe plicatilis) is a tree aloe endemic to mountain fynbos in the Boland, south-western Cape. The viability and germination behaviour of K. plicatilis seeds were assessed for seeds stored for four and nine months at − 80 °C, 4 °C, 25 °C and under ambient conditions in a laboratory. Seeds were germinated under controlled conditions and germination rates and percentages determined. Ungerminated seeds were tested for viability using tetrazolium salt. Seed viability was not significantly reduced during storage. Seeds stored at − 80 °C for four and nine months exhibited the fastest germination rate overall (both 5.9 ± 0.3 weeks, mean ± S.E.), and slowest was for seeds stored under ambient conditions for four and nine months (both 7.8 ± 0.4 weeks). All seed lots showed similar percentage germination after four months of storage (78.0–90.4%). The highest percentage germination overall was for seeds stored at − 80 °C for four months (90.4%) and the lowest was for seeds kept at 4 °C and − 80 °C for nine months (39.2 and 39.6%, respectively). Respective percentage viability for ungerminated seeds in these two treatments was 82% and 87%, respectively, indicating the induction of secondary dormancy. Induced dormancy triggered by protracted cold temperatures may be an adaptation that enables seeds to survive prolonged extreme conditions that are unfavourable for germination. Further research on the long-term storage of aloe seeds would be beneficial for developing long-term seed storage and germination testing protocols for ex situ conservation.     

Oestrus cycle characteristics and prediction of ovulation in Catalonian jennies

The Catalonian donkey breed is in danger of extinction, and much needs to be learned about the reproductive features of its females if breeding and conservation programmes are to be successful. This study reports the oestrous behaviour, oestrus cycle characteristics and dynamic ovarian events witnessed during 50 oestrous cycles (involving 106 ovulations) in 10 Catalonian jennies between March 2002 and January 2005. These jennies were teased, palpated transrectally and examined by ultrasound using a 5 MHz linear transducer—daily during oestrus and every other day during dioestrus. Predictors of ovulation were sought among the variables recorded.The most evident signs of oestrus were mouth clapping (the frequent vertical opening and closing of the mouth with ears depressed against the extended neck) and occasional urinating and winking of the vulval lips (homotypical behaviour). Interactions between jennies in oestrus were also recorded, including mounting, herding/chasing, the Flehmen response, and vocalization (heterotypical behaviour).Nine jennies ovulated regularly throughout the year; one had two anovulatory periods (54 and 35 days). The length of the oestrus cycle was 24.90 ± 0.26 days, with oestrus itself lasting 5.64 ± 0.20 days (mean ± S.E.M.) and dioestrus 19.83 ± 0.36 days. The incidence of single, double and triple ovulations was 55.66% (n = 59), 42.45% (n = 45) and 1.89% (n = 2), respectively. No significant difference was seen in the number of ovulations involving the left and right ovaries (52.63% [n = 70] compared to 47.37% [n = 63] respectively; P > 0.05). The mean interval between double ovulation was 1.44 ± 3.98 days. The mean diameter of the preovulatory follicle at day −1 was 44.9 ± 0.5 mm; the mean growth rate over the 5 days before ovulation was 3.7 mm/day.Data on preovulatory changes in oestrous behaviour, follicle size, follicle texture, the echographic appearance of the follicle and uterus, and uterine tone were subjected to stepwise logistic regression analysis to detect predictors of ovulation. The logit function showed the best predictors to be follicle size, follicular texture and oestrous behaviour. Certain combinations of these three variables allow the prediction of ovulation within 24 h with a probability of >75%.     

The effect of internal capacitance on power quality and energy efficiency in a tubular microbial fuel cell

The pseudo-capacitive behaviour of a high surface area carbon veil electrode in a tubular microbial fuel cell (MFC) was investigated as a mechanism to enhance power quality and energy efficiency. Accumulated charge and energy from the anodic biofilm after prolonged open circuit times (1–120 min) were compared against equivalent periods of steady state loading (R = 100–3000 Ω). A significant difference in the amount of accumulated charge with different loads was observed, resulting in 1.051 C (R = 100 Ω) compared to 0.006 C (R = 3 kΩ). The automated application of short open and closed circuit (0.5–10 s) cycles resulted in an increase of power/current production (closed circuit alone), but presented lower efficiency considering entire open and closed period. The cumulative charge on the carbon veil electrode with biofilm was 39,807 C m⁻² at 100 Ω. Electrochemical Impedance Spectroscopy (EIS) showed that the Helmholtz layer presented a double layer capacitance of more than ten times the biofilm on electrode. The results indicate that the capacitive behaviour could be utilized to increase the power quality, i.e. its availability/applicability with respect to the operation of low power consuming devices.     

Feeding,resting and social behaviour in ewes housed in two different group sizes

The aim of this experiment was to investigate the effects of increased group size on eating- and resting behaviour, aggression and feed intake in housed ewes. During an initial period of 14 days 36 adult (2–6 years old) ewes of the domestic Norwegian Dala breed were divided into four groups of 9. In the second period (14 days), these ewes were merged into one group of 36 ewes. This experiment was repeated with a second batch of ewes, but this time starting with a group of 36 individuals in the first period, then splitting them up into four groups of 9 ewes in the second period. From 24 h video recordings we scored activity behaviours using instantaneous sampling every 10 min. Aggressive interactions were continuously observed the first 10 min every hour during the 24 h (4 h in total). A mixed statistical procedure with group size, day, batch and the interactions between them were included as fixed effects, whereas individual and group were specified as random effects.Ewes in large groups (36) had a larger variation in lying time at day one (P < 0.01), less synchronized lying (P < 0.05) and eating behaviour (P < 0.01), and spent less time queuing at the feed barrier (P < 0.001) compared to in the small group size (9). There were no effects of group size on aggressive interactions or feed intake.In conclusion, a larger group size decreased synchrony in resting and feeding behaviour and reduced the time spent queuing in front of the feed barrier. It is possible that the aggression level in sheep is more sensitive to changes in space allowance than to changes in group size per se.     

Optimization of a rapid microwave-assisted extraction method for the simultaneous determination of opiates,cocaine and their metabolites in human hair

A rapid and cleanup-free microwave-assisted extraction (MAE) method is proposed for the simultaneous extraction of six illegal drugs of abuse – cocaine, benzoylecgonine (BZE), cocaethylene (CCE), morphine, 6-monoacethylmorphine (6AM) and codeine – from human hair samples. The analytes were determined using high performance liquid chromatography (HPLC) with photodiode array UV detection. The influence of several variables on the efficiency of the MAE procedure was investigated in detail by a multi-objective optimization approach based on a hybrid experimental design (17 experiments) and desirability functions. Six drugs were successfully extracted from human hair with recoveries close to 100% and good reproducibility (<3.6% RSD) under the optimal MAE conditions: 11 mL dichloromethane (DCM) extraction solvent, 60 °C extraction temperature, 9 min extraction time and 0.5 mL of methanol (MeOH) added to 50 mg of the hair sample in the extraction vessels. Limits of quantification of 0.2 ng mg^?1 were found for the studied compounds. A comparison of sample preparation procedures, including MAE, enzymatic digestion and digestion by aqueous acids, was also conducted. The results indicated that the global behaviour of sample procedures provided similar satisfactory recoveries ranging from 86 to 100%. Indeed, the MAE procedure resulted in a reduction of extraction time by 100-fold and the elimination of cleanup steps. Slightly higher recoveries of morphine, 6AM, BZE and CCE, at 1 ng mg^?1 concentration level and cocaine at 40 ng mg^?1 concentration level, were achieved using MAE. Lastly, the proposed MAE method was applied to several human hair samples from multidrug abusers.     

The use of a mirror reduces isolation stress in horses being transported by trailer

Horse trailers are a common form of transportation for horses and ponies and often require the animal to travel alone or with a single companion. The current study investigated the effect of transporting horses alone, in company or with an acrylic safety mirror (measuring 81 cm × 61.5 cm) that provided surrogate companionship. The behavioural and physiological responses of 12 mature horses during a 30-min journey by trailer under the three treatments were compared. Behaviours (vocalisation, eating, head-tossing, pawing, and head-turning) were recorded. In order to assess circulatory changes that occur as part of the response to transport, heart rate (HR), rectal (T_r) and ear-pinna (T_p) temperatures were recorded. When travelling with a live companion significantly less time was spent vocalising (p < 0.001), head-turning (p < 0.001), head-tossing (p < 0.01) and pawing (p < 0.01); eating behaviour increased (p < 0.05). Physiological responses (increases in HR and T_r and decreases in T_p) were also significantly reduced when travelling with a live companion (p < 0.01). Travelling with the mirror did not significantly affect physiological responses compared with travelling alone, but the rise in T_r and fall in T_p was reduced (p = 0.052 and p = 0.051, respectively) and can be considered a trend. When travelling with a mirror significantly less time was spent turning the head (p < 0.01), vocalising (p < 0.05) and head-tossing (p < 0.05); eating behaviour increased (p < 0.05). The only significant difference between travelling with a live companion and a mirror was that the time spent turning the head round was less with a live companion (p < 0.05). The provision of surrogate companionship in the form of a mirror was found to be preferable to travel alone, but where possible a live companion is recommended. Isolation during transportation was found to suppress feeding behaviour. Although peripheral blood flow (T_p) has been used to assess transport stress in other species it has not previously been used in the horse. Further evaluation of this non-invasive measure is now required.     

The effects of muscle architectural change with a pre-motion silent period on the subsequent muscular output during rapid voluntary movement

This study aimed to determine the characteristics of the in vivo behaviour of human muscle architecture during a pre-motion silent period (PMSP) using ultrasonography. Subjects were requested to perform rapid knee extension with vertical jumping. Electromyographic signals were recorded from the vastus lateralis (VL), vastus medialis, and biceps femoris muscles. Ultrasonic images were recorded from the VL. We found that the cross point between the fascicle and deep aponeurosis in the VL moved to the distal side before the rapid vertical jumps with PMSP. This cross point movement with PMSP was of low amplitude (mean: 1.0 ± 0.3 mm) and velocity (22.2 ± 6.1 mm/s). The amplitude and velocity of the cross point movement were significantly positively related to the angular peak velocity of knee extensor during rapid vertical jumping in trials with PMSP. These results suggest that although low levels of pre-movement muscle architectural change with PMSP may be the result of muscle relaxation behaviour rather than the result of muscle stretching behaviour, this pre-movement effect can influence subsequent muscular performance during a rapid voluntary movement. PMSP may allow pre-movement muscle architectural change to generate a better muscular condition to increase neural activation during the subsequent rapid voluntary contraction.     

The effect of breed and housing system on dairy cow feeding and lying behaviour

In Ireland there is growing interest in managing dairy cows on out-wintering pads (OWPs) during the winter, as a low cost alternative to housing indoors. This study investigated feeding and lying behaviour of two breeds of dairy cow (Holstein-Friesian and Norwegian Red) at pasture (PAS) and in winter (WIN) confinement. Cows (n = 36) were managed as one herd while lactating at PAS, then dried off on entering WIN on 17 November 2005 and assigned to one of the three treatments using a randomised complete block design: (1) indoor matted cubicles [IC], (2) unsheltered OWP [UP] and (3) sheltered OWP [CP] (feed system = concrete feedface). Feeding behaviour was recorded for 1 × 24 h period during PAS and WIN using IGER grazing behaviour recorders. Standing/lying was recorded every 5 min for 2 × 24 h periods at PAS and 1 × 24 h period during WIN using modified voltage dataloggers (Tinytag Plus, Chichester, UK). Although not compared statistically, cows spent more time feeding at PAS (530.7 ± 69.66 min/day) than in any winter confinement system (UP, IC, CP = 453.9 ± 37.36, IC = 462.7 ± 37.31 and CP = 505.9 ± 37.36 min/day). In contrast, cows spent more time lying during the winter period (UP, IC, CP = 11.7 ± 0.45, IC = 10.8 ± 0.39 and CP = 11.0 ± 0.39 h/day) than when at PAS (9.4 h/day). Holstein-Friesian cows had a higher bite rate and fewer mastications while feeding, than Norwegian Red cows (P < 0.05 for both). Shorter feeding times at WIN were likely primarily due to lower metabolic requirements associated with the dry period, although the higher fibre content of the silage may also have contributed. The increase in ruminating times between PAS and WIN is likely also due to the higher fibre content of silage than grass. Differences in feeding behaviour between breeds may be indicative of reduced motivation to ingest food quickly. This may be an indirect consequence of a selection index that has led to improvements in body condition and self-maintenance. The similarity in lying and feeding times during WIN indicates that during these winter weather conditions the feeding and lying behaviour of cows on unsheltered OWPs was not inhibited compared with sheltered cows.     

A reduced core to skin temperature gradient,not a critical core temperature,affects aerobic capacity in the heat

The purpose of this study was to determine the impact of the core to skin temperature gradient during incremental running to volitional fatigue across varying environmental conditions. A secondary aim was to determine if a “critical” core temperature would dictate volitional fatigue during running in the heat. 60 participants (n=49 male, n=11 female; 24±5 yrs, 177±11 cm, 75±13 kg) completed the study. Participants were uniformly stratified into a specific exercise temperature group (18 °C, 26 °C, 34 °C, or 42 °C) based on a 3-mile run performance. Participants were equipped with core and chest skin temperature sensors and a heart rate monitor, entered an environmental chamber (18 °C, 26 °C, 34 °C, or 42 °C), and rested in the seated position for 10 min before performing a walk/run to volitional exhaustion. Initial treadmill speed was 3.2 km h⁻¹ with a 0% grade. Every 3 min, starting with speed, speed and grade increased in an alternating pattern (speed increased by 0.805 km h⁻¹, grade increased by 0.5%). Time to volitional fatigue was longer for the 18 °C and 26 °C group compared to the 42 °C group, (58.1±9.3 and 62.6±6.5 min vs. 51.3±8.3 min, respectively, p<0.05). At the half-way point and finish, the core to skin gradient for the 18 °C and 26 °C groups was larger compared to 42 °C group (halfway: 2.6±0.7 and 2.0±0.6 vs. 1.3±0.5 for the 18 °C, 26 °C and 42 °C groups, respectively; finish: 3.3±0.7 and 3.5±1.1 vs. 2.1±0.9 for the 26 °C, 34 °C, and 42 °C groups, respectively, p<0.05). Sweat rate was lower in the 18 °C group compared to the 26 °C, 34 °C, and 42 °C groups, 3.6±1.3 vs. 7.2±3.0, 7.1±2.0, and 7.6±1.7 g m⁻² min⁻¹, respectively, p<0.05. There were no group differences in core temperature and heart rate response during the exercise trials. The current data demonstrate a 13% and 22% longer run time to exhaustion for the 18 °C and 26 °C group, respectively, compared to the 42 °C group despite no differences in beginning and ending core temperatures or baseline 3-mile run time. This capacity difference appears to result from a magnified core to skin gradient via an environmental temperature advantageous to convective heat loss, and in part from an increased sweat rate.     

Unusual change in activity pattern at cool temperature in a reptile (Sphenodon punctatus)

Animals that can be active both during day and night offer unique opportunities to identify factors that influence activity pattern. By experimental manipulations of temperatures under constant photoperiod, we aimed to determine if emergence, activity and thermoregulatory behaviour of juvenile tuatara (Sphenodon punctatus) varied at different temperatures (20 °C, 12 °C and 5 °C). To help clarify its activity pattern, we compared tuatara with two lizard species endemic of the South Island of New Zealand for which activity pattern is known and clearly defined: the nocturnal common gecko Woodworthia “Otago/Southland” and the diurnal McCann׳s skink Oligosoma maccanni. Tuatara showed similar responses to both species of lizards. Similar to the diurnal skinks, tuatara emerged quickly at 20 °C and 12 °C while nocturnal geckos took more time to emerge. Like nocturnal geckos, tuatara continued to be active at 5 °C, but only during the day. Interestingly, tuatara shifted from diurno-nocturnal activity at 20 °C and 12 °C to being strictly diurnal at 5 °C. We suggest that this temperature-dependent strategy maximises their survival during cold periods.     

Feeding enrichment in an opportunistic carnivore: The red fox

In captive carnivores, species-specific behaviour is often restricted by inadequate feeding regimens. Feeding live prey is not feasible in most places and food delivery is often highly predictable in space and time which is considerably different from the situation in the wild. As a result, captive carnivores are often inactive, show little behavioural diversity and are prone to behavioural problems such as stereotypic pacing. Using artificial feeding devices to substitute for natural food resources is a way to address these problems. In a group of four red fox (Vulpes vulpes), we compared a conventional feeding method to four different methods through the use of feeding enrichment that were based on natural foraging strategies of opportunistic carnivores. Feeding enrichments consisted of electronic feeders delivering food unpredictable in time which were successively combined with one of the three additional treatments: a self-service food box (allowing control over access to food), manually scattering food (unpredictable in space), and an electronic dispenser delivering food unpredictably both in space and time. The aim of administering feeding enrichment in this study was to stimulate appetitive (food searching) behaviour and to increase time spent in feeding. Compared to conventional feeding, diversity of behaviour and overall activity were significantly enhanced in the presence of electronic feeders in all four foxes (EF > CON1 = CON2, EF + SF > CON1 = CON2, EF + MS > CON1 = CON2, EF + ED > CON1 = CON2). Behavioural diversity was highest when the foxes had control over access to food (EF + SF), while the manual scattering of food (EF + MS) and the electronic dispenser (EF + ED) enhanced food searching behaviour. These results indicate that in opportunistic carnivores natural foraging and feeding behaviour can be stimulated by simple feeding enrichment strategies, and that foraging behaviour is stimulated most when food delivery is unpredictable both in space and time.     

Direct effects of incubation temperature on morphology,thermoregulatory behaviour and locomotor performance in jacky dragons (Amphibolurus muricatus)

Incubation temperature is one of the most studied factors driving phenotypic plasticity in oviparous reptiles. We examined how incubation temperature influenced hatchling morphology, thermal preference and temperature-dependent running speed in the small Australian agamid lizard Amphibolurus muricatus. Hatchlings incubated at 32 °C grew more slowly than those incubated at 25 and 28 °C during their first month after hatching, and tended to be smaller at one month. These differences were no longer significant by three months of age due to selective mortality of the smallest hatchlings. The cooler incubation treatments (25 °C and 28 °C) produced lizards that had deeper and wider heads. Hatchlings from 28 °C had cooler and more stable temperature preferences, and also had lower body temperatures during a 2-h thermoregulatory behaviour trial. Locomotor performance was enhanced at higher body temperatures, but incubation temperature had no measurable effect either independently or in interaction with body temperature. Our study demonstrates that incubation temperature has direct effects on morphology and thermoregulatory behaviour that appears to be independent of any size-dependent effects. We postulate a mechanistic link between these two effects.     

Oestrus in the Julia Creek dunnart (Sminthopsis douglasi) is associated with wheel running behaviour but not necessarily changes in body weight,food consumption or pouch morphology

The Julia Creek dunnart (Sminthopsis douglasi) is an endangered carnivorous marsupial belonging to the family Dasyuridae. This study investigated the oestrous cycle of this species in terms of its reproductive physiology and behaviour to explore more efficient methods of oestrus detection. Ten sexually mature captive female dunnarts were monitored daily at David Fleay Wildlife Park, Burleigh Heads, Australia, from mid September to late December 2006 for changes in urogenital cytology within the urine (0, 1+, 2+ and 3+), running wheel activity, body weight, uneaten food, faecal steroid metabolites (progesterone and oestradiol) and pouch development. Periods of increased running wheel activity were associated (p = 0.004) with an increase in the proportion of cornified urogenital epithelial cells found in the urine; periods of decreasing weight (p < 0.001) and uneaten food (p < 0.001) were also associated with changes in urogenital cytology but not to the point where they would be useful for oestrus detection. Between 60.3% and 92.0% of peak distances (confidence interval 95%) occurred when the epithelial cell index was 2+ or 3+. Only 15.5–37.5% of peak weights (CI: 95%) and 28.1–49.9% of incidences of uneaten food (CI: 95%) occurred when the epithelial cell index was 2+ or 3+. There was no significant difference in the mean length of the oestrous cycle when measured by urogenital cytology (mean ± SD: 25.0 ± 5.7 days; n = 20) or peak distance travelled (mean ± SD: 25.4 ± 5.7 days; n = 20). Changes in the concentration of oestradiol metabolites in Julia Creek Dunnart faeces were not useful in characterising the oestrous cycle. Wheel running activity declined markedly with increased faecal progestagen concentration. The majority of the pouch variables examined showed maximum development during the inter-oestrus period but as there was considerable variation between animals, the pouch was not considered a useful index of oestrus.     

Ambient temperature and humidity modulate the behavioural thermoregulation of a small arboreal mammal (Callicebus bernhardi)

Maintaining a constant body temperature is critical to the proper functioning of metabolic reactions. Behavioural thermoregulation strategies may minimize the cost of energetic balance when an animal is outside its thermoneutral zone. We investigated whether ambient temperature and relative air humidity influence the use of behavioural strategies by a group of Prince Bernhard's titi monkeys (Callicebus bernhardi) living in a forest fragment. We monitored a social group composed of four individuals (an adult couple and two juveniles) for 1010 h from March to September 2015. We used the instantaneous scan sampling method to record the body posture, the microhabitat, and the occurrence of huddling with group mate(s) when animals were resting. We recorded ambient temperature and relative humidity in the shade every 10 min with a data logger hanging at a height of approximately 5 m. Daytime temperature ranged from 18.5 °C to 38.5 °C and relative humidity ranged from 21% to 97%. Titi monkeys avoided sunny places at higher temperatures, especially above 31 °C. Minimum night temperature did not influence the choice of resting microhabitats during the first hour after sunrise. Sitting was the major resting posture during the day (62%). Titi monkeys increased the use of heat-dissipating postures at ambient temperatures >27 °C. In addition, an increase in relative humidity increased the use of these postures at 26 °C, 27 °C, 29 °C and 33 °C, but caused a decrease at 24 °C. On the other hand, the ambient temperature did not influence the occurrence of huddling. We conclude that microhabitat choice and postural behaviour are important for titi monkeys to prevent overheating and suggest that these behavioural adjustments might also be critical for other tropical arboreal mammals.     

GLP-1(32–36)amide,a novel pentapeptide cleavage product of GLP-1, modulates whole body glucose metabolism in dogs

We have previously demonstrated in human subjects who under euglycemic clamp conditions GLP-1(9–36)amide infusions inhibit endogenous glucose production without substantial insulinotropic effects. An earlier report indicates that GLP-1(9–36)amide is cleaved to a nonapeptide, GLP-1(28–36)amide and a pentapeptide GLP-1(32–36)amide (LVKGR amide). Here we study the effects of the pentapeptide on whole body glucose disposal during hyperglycemic clamp studies. Five dogs underwent indwelling catheterizations. Following recovery, the dogs underwent a 180 min hyperglycemic clamp (basal glucose +98 mg/dl) in a cross-over design. Saline or pentapeptide (30 pmol kg⁻¹ min⁻¹) was infused during the last 120 min after commencement of the hyperglycemic clamp in a primed continuous manner. During the last 30 min of the pentapeptide infusion, glucose utilization (M) significantly increased to 21.4 ± 2.9 mg kg⁻¹ min⁻¹compared to M of 14.3 ± 1.1 mg kg⁻¹ min⁻¹ during the saline infusion (P = 0.026, paired t-test; P = 0.062, Mann–Whitney U test). During this interval, no significant differences in insulin (26.6 ± 3.2 vs. 23.7 ± 2.5 μU/ml, P = NS) or glucagon secretion (34.0 ± 2.1 vs. 31.7 ± 1.8 pg/ml, P = NS) were observed. These findings demonstrate that under hyperglycemic clamp studies the pentapeptide modulates glucose metabolism by a stimulation of whole-body glucose disposal. Further, the findings suggest that the metabolic benefits previously observed during GLP-1(9–36)amide infusions in humans might be due, at least in part, to the metabolic effects of the pentapeptide that is cleaved from the pro-peptide, GLP-1(9–36)amide in the circulation.     

Active compounds from a diverse library of triazolothiadiazole and triazolothiadiazine scaffolds: Synthesis,crystal structure determination,cytotoxicity, cholinesterase inhibitory activity,and binding mode analysis

In an effort to identify novel cholinesterase candidates for the treatment of Alzheimer’s disease (AD), a diverse array of potentially bioactive compounds including triazolothiadiazoles (4a–h and 5a–f) and triazolothiadiazines (6a–h) was obtained in good yields through the cyclocondensation reaction of 4-amino-5-(pyridin-3-yl)-4H-1,2,4-triazole-3-thiol (3) with various substituted aryl/heteroaryl/aryloxy acids and phenacyl bromides, respectively. The structures of newly prepared compounds were confirmed by IR, ¹H and ¹³C NMR spectroscopy and, in case of 4a, by single crystal X-ray diffraction analysis. The purity of the synthesized compounds was ascertained by elemental analysis. The newly synthesized conjugated heterocycles were screened for cholinesterase inhibitory activity against electric eel acetylcholinesterase (EeAChE) and horse serum butyrylcholinesterase (hBChE). Among the evaluated hybrids, several compounds were identified as potent inhibitors. Compounds 5b and 5d were most active with an IC₅₀ value of 3.09 ± 0.154 and 11.3 ± 0.267 μM, respectively, against acetylcholinesterase, whereas 5b, 6a and 6g were most potent against butyrylcholinesterase, with an IC₅₀ of 0.585 ± 0.154, 0.781 ± 0.213, and 1.09 ± 0.156 μM, respectively, compared to neostigmine and donepezil as standard drugs. The synthesized heteroaromatic compounds were also tested for their cytotoxic potential against lung carcinoma (H157) and vero cell lines. Among them, compound 6h exhibited highest antiproliferative activity against H157 cell lines, with IC₅₀ value of 0.96 ± 0.43 μM at 1 mM concentration as compared to vincristine (IC₅₀ = 1.03 ± 0.04 μM), standard drug used in this study.     

Liquid chromatography/tandem mass spectrometry method for the simultaneous determination of vardenafil and its major metabolite,N-desethylvardenafil,in human plasma: Application to a pharmacokinetic study

A rapid and sensitive LC–MS/MS method for the determination of vardenafil and its major metabolite, N-desethylvardenafil, in human plasma using sildenafil as an internal standard was developed and validated. The analytes were extracted from 0.25-mL aliquots of human plasma by liquid–liquid extraction, using 1 mL of ethyl acetate. Chromatographic separation was carried on a Luna C₁₈ column (50 mm × 2.0 mm, 3 μm) at 40 °C, with an isocratic mobile phase consisting of 10 mM ammonium acetate (pH 5.0) and acetonitrile (10:90, v/v), a flow rate of 0.2 mL/min, and a total run time of 2 min. Detection and quantification were performed using a mass spectrometer in the selected reaction-monitoring mode with positive electrospray ionization at m/z 489.1 → 151.2 for vardenafil, m/z 460.9 → 151.2 for N-desethylvardenafil, and m/z 475.3 → 100.1 for the internal standard (IS), respectively. This assay was linear over a concentration range of 0.5–200 ng/mL with a lower limit of quantification of 0.5 ng/mL for both vardenafil and N-desethylvardenafil. The coefficient of variation for the assay precision was <13.6%, and the accuracy was >93.1%. This method was successfully applied to a pharmacokinetic study after oral administration of vardenafil 20 mg tablet in Korean healthy male volunteers.     

Comparative diel oxygen cycles preceding and during a Karenia bloom in Sarasota Bay,Florida, USA

The diel change in dissolved oxygen concentrations were recorded with an automated incubator containing a pulsed oxygen sensor in Sarasota Bay, Florida. The deployments occurred during a ‘pre-bloom’ period in May to June 2006, and during a harmful algal bloom dominated by Karenia brevis in September 2006. The diurnal (daylight) increase in dissolved oxygen concentrations varied from 16 to 104 μmol O₂ l⁻¹ with the corresponding nocturnal decrease in oxygen varying from 16 to 77 μmol O₂ l⁻¹. Nocturnal respiration consumed 42–113% of the diurnal net oxygen production with the minimum and maximum during the pre-bloom period. Hourly production rates closely followed fluctuations in irradiance with maximum rates in the late morning. Hourly oxygen utilization rates (community respiration) at night were highest during the first few hours after sunset.