The volume of distribution (Vd) is 1.1 L/kg both in neonates and adults. Midazolam is hydroxylated by CYP3A4 and CYP3A5; the activities of these enzymes surge in the liver in the first weeks of life and thus the metabolic rate of midazolam is lower in neonates than in adults. Midazolam acts as a sedative, as an antiepileptic, for those infants who are refractory to standard antiepileptic therapy, and as an anaesthetic. Information of midazolam as an anaesthetic in infants are very little. Midazolam is usually administered intravenously; when minimal sedation is required, intranasal administration of midazolam is employed. Disease affects the pharmacokinetics of midazolam in neonates; multiple organ failure reduces the Cl of midazolam and mechanical ventilation prolongs the t1/2 of this drug. ECMO therapy increases t1/2, Cl, and Vd of midazolam several times. The adverse effects of midazolam in neonates are scarce: pain, tenderness, and thrombophlebitis may occur. Respiratory depression and hypotension appear in a limited percentage of infants following intravenous infusion of midazolam. In conclusion, midazolam is a safe and effective drug which is employed as a sedative, as antiepileptic agent, for infants who are refractory to standard antiepileptic therapy, and as an anaesthetic.Midazolam is a benzodiazepine with rapid onset of action and short duration of effect. In healthy neonates the half-life ( Midazolam is a short-acting benzodiazepine with rapid onset of action. It has anxiolytic, muscle relaxant, and anticonvulsant activity, now most widely used to generate anterograde amnesia and to stop prolonged seizures in children . MidazolA receptors are responsible for most inhibitory neurotransmission in the central nervous system. Benzodiazepines act at GABAA receptors by binding directly to a specific site that is distinct from that of GABA binding [Midazolam exerts most of its effects by interacting with inhibitory neurotransmitter receptors directly activated by GABA. GABA binding . Midazol binding .t1/2) of midazolam is 1.9 hours, which is 22-fold shorter than that of diazepam [t1/2 of midazolam is shorter in neonates; it is about from 4 to 6 hours. It is quite variable and may be up to 22 hours in premature infants [In adults, the half-life  or with the sedative-hypnotic midazolam in ventilated neonates. Endotracheal intubation and mechanical ventilation are major components of routine intensive care for very low birthweight newborn infants and sick fullterm newborn infants . Sedatioμg/kg per min, with an average of 4 μg/kg per min, terminated the seizures in 32 infants whose seizures could not be controlled by diazepam, phenytoin, or phenobarbital. Castro Conte et al. [μg/kg per min, increasing by 0.5 to 1 μg/kg every 2 min, until a maximum of 18 μg/kg per min) controlled the seizures in 13 neonates refractory to the treatment with phenobarbital and phenytoin.A useful action of midazolam is the management of neonatal seizures refractory to conventional treatments. Hu et al.  reportede et al.  found thTroponin T is a prognostic indicator of postoperative recovery. Midazolam, propofol, or sevoflurane had similar efficacy in the production of troponin T in infants who underwent cardiac surgery .The information on the effects of midazolam and on the fate of this drug in neonates has been published in different journals during the last thirty years and it is scattered. The aims of this article are (1) to gather together and (2) to review the published data on (a) the metabolism and (b) the pharmacokinetics of midazolam in neonates, (c) the therapeutic effects, and finally (d) the adverse effects of midazolam in neonates. The main objective of this work is (e) to provide neonatologists with a tool that embraces all aspects of the clinical pharmacology of midazolam in neonates. Little is known about the effects and the fate of the drugs administered to neonates. This review was written to help the neonatologists, hoping that they will find some useful information and thus some advantages for their work, from reading the present paper.The bibliographic search was performed electronically using PubMed and EMBASE databases as search engines; September 2013 was the cutoff point. The following key words: “midazolam neonate,” “midazolam mechanism of action,” “midazolam therapy neonate,” “midazolam pharmacokinetics neonate,” “midazolam metabolism neonate,” “CYP3A4 midazolam neonate,” “CYP3A5 midazolam neonate,” “oral administration midazolam neonate,” “intranasal administration of midazolam neonates,” “midazolam seizures,” “midazolam anaesthesia neonate,” and “midazolam adverse effects neonate,” were used. The bibliography of each article was read carefully, and the selected articles were examined. In addition, the books NEOFAX: a Manual Used in the Neonatal Care, by Young and Mangum [μg/kg of midazolam over at least 5 min. Midazolam may also be administered intramuscularly. Repeat as required, usually Q2 to 4 hours. Dosage requirements are decreased by concurrent use of narcotics. For continuous intravenous infusion, give a 10 to 60 μg/kg midazolam per hour. Dosage of this drug may need to be increased after several days of therapy because of development of tolerance and/or increase of Cl. For Intranasal administration, give a 200 to 300 μg/kg midazolam per dose using 5 mg/mL injectable form. For sublingual administration, give a 0.2 mg/kg per dose of midazolam using 5 mg/mL injectable form mixed with small amounts of flavoured syrup. For Oral administration, give 250 μg per dose. For Anticonvulsant therapy; give intravenously a loading dose of 150 μg/kg over at least 5 min followed by a maintenance infusion dose of 60 to 400 μg/kg per hour (1 to 7 μg/kg per min).For sedation, Young and Neofax  suggest Development has a profound effect on the expression of phase I enzymes such as the cytochromes P450 (CYPs). Distinct patterns of isoform specific development CYP expression have been observed postnatally. Although the CYP content in the fetal liver equals about 30 to 60% of adult values , isoformIn adults, midazolam is metabolized rapidly ; the recPlasma Cl of midazolam is widely used as an in vivo surrogate measurement of CYP3A4 and CYP3A5 activities , 45. Cl Ince et al.  developeμg/kg) was administered intravenously (n = 15) or orally (n = 7). After intravenous administration, the median percentage of the dose excreted into the urine, over 6-hour intervals, was 0.44% (midazolam), 0.04% (1-hydroxymidazolam), and 1.57% (1-hydroxymidazolam glucuronide). After oral administration, the median percentage of the dose excreted into the urine was 0.11% (midazolam), 0.02% (1-hydroxymidazolam), and 1.69% (1-hydroxymidazolam glucuronide). The glucuronidation of 1-hydroxymidazolam is immature in preterm infants less than 2 weeks of postnatal age.The ontogeny of midazolam glucuronidation was studied in 22 preterm infants . MidazolThe Cl of midazolam was measured in 24 preterm infants and the mean was 1.8 (mL/kg per min) . In chilThe published literature on the pharmacokinetics of midazolam in neonates is scarce and the available information is heterogeneous because the work was done in healthy neonates , in critThe information on the pharmacokinetics of midazolam summarised in this review is organised on the neonate health status, and it is reported in (a) healthy neonates, (b) in critically ill neonates (c), and in infants undergoing extracorporeal membrane oxygenation (ECMO) and finally (d) the population pharmacokinetics of midazolam are reviewed in neonates. The pharmacokinetic parameters of midazolam are summarised in μg/kg) were studied in 24 healthy preterm infants, with a gestational age between 26 and 34 weeks [t1/2 was 6.3 hours (range: 2.6 to 17.7 hours), Cl was 1.8 mL/kg per min (range 0.7 to 6.7 mL/kg per min), Vd was 1.1 L/kg (range: 0.4 to 4.2 L/kg), and AUC0–∞ was 971 ng·h/mL (range: 248 to 2353 ng·h/mL). In the text, these values are referred to as the “normal values” or the “normal estimates.”The pharmacokinetics of intravenous midazolam  and Tmax was 6 hours (range: 1 to 12 hours). The median 1-hydroxymidazolam AUCt0– to midazolam AUCt0– ratio was low (0.09) and showed a large interindividual variation (range: <0.001 to 1). There were no relationships between 1-hydroxymidazolam pharmacokinetic parameters and midazolam Cmax, AUC, and t1/2. No 4-hydroxymidazolam could be detected.No significant relationships were detected between age  and midazolam Cl, Vd, and t1/2 was 73.6 weeks. Predicted Cl changes with age, based on this model, were in close agreement with physiologically based pharmacokinetic models. Previously published pharmacokinetic parameters can be used to develop maturation models that address gaps in current knowledge regarding the influence of age on a drug's disposition. If midazolam sedation target concentration of 100 ng/mL, similar to that given to adults, is assumed, we might anticipate steady-state infusion rates of 14 μg/kg per hour in neonates, 50 μg/kg per hour in a 1-year-old infant, 60 μg/kg per hour in a 5-year-old infant, and 50 μg/kg per hour in a 12-year-old child. Age-related pharmacodynamic differences that will affect dose and the impact of active metabolites on response have not yet been quantified.Anderson and Larsson  describet1/2 was 9.8 hours, after a 200 μg/kg bolus of midazolam, and 12 hours, after a constant infusion of 60 μg/kg per hour of midazolam [t1/2 is 6.3 hours. These estimates are 155% and 190%, respectively, of the normal values [Disease reduces the midazolam elimination rate in neonates. In critically ill neonates, undergoing mechanical ventilation, idazolam . In norml values .μmol/L, received an intravenous bolus of 100 μg/kg midazolam. t1/2, Cl, and Vd of midazolam of this girl were 6.3-fold longer, 2.6-fold greater, and 14.5-fold larger, respectively, [A 15.1-year-old girl, suffering from renal failure, with malignant hypertension and with a creatinine concentration of 691 ctively,  than thectively, . The patctively, .t1/2 and a Cl of midazolam of 3.8-fold longer and 1.8-fold greater, respectively, [A 1.1-year-old boy, suffering from pneumonia, received the CYP3A4 inhibitor erythromycin and the CYP3A4 substrate fentanyl. He had a ctively,  than thectively, .μg/kg per hour (range: 50 to 270 μg/kg per hour). The pharmacokinetic parameters of midazolam are summarised in P = 0.035). A significant negative correlation was found between disease severity and midazolam Cl corrected for body weight . These results suggest that increased disease severity is associated with reduced midazolam Cl. Likely, these results reflect a reduction of CYP3A activity.Vet et al.  determinAlternative explanations could be the altered protein binding, midazolam being highly bound to plasma proteins , or, leμg/kg per hour; the infusion lasted for 60.0 ± 23.3 hours. In these patients, t1/2 estimate was double the normal value [μg/mL; in two cases, the patients received fentanyl before midazolam. The metabolic ratio of midazolam to 1-hydroxymidazolam, was 16.1 ± 21.2 (n = 11) at 48 hours after the administration. Various drugs were coadministered with midazolam during the study; they were fentanyl (n = 7), antibiotics, in most cases betalactams and aminoglycosides (n = 13), albumin (n = 5), and vasopressin drugs (n = 3). Jacqz-Aigrain et al. [t1/2.The pharmacokinetics of midazolam was studied in 15 preterm infants undergoing mechanical ventilation for respiratory distress syndrome . Midazolal value . Hypotenal value . Mean syn et al.  concluden et al. , can inhμg/kg of midazolam [t1/2, Cl, and Vd were 6.5 ± 2.3 hours, 2.0 ± 1.2 mL/kg per min, and 0.9 ± 0.3 L/kg, respectively. The kinetic parameters in these infants were similar to those of normal values [Ten critically ill preterm infants, with a postnatal age of 2 to 5 days, received a single intravenous bolus dose of 200 idazolam . Midazoll values .μg/kg followed by an intravenous infusion of either 50 μg/kg per hour, for postoperative monitoring, or 100 μg/kg per hour for conscious sedation). In children, a large variability in the pharmacokinetic parameters of midazolam was observed. Midazolam is a substrate for CYP3A4 and CYP3A5 and the expression of these enzymes considerably varies in neonates [Ince et al.  performeneonates , 10, 11.The wide variability of midazolam pharmacokinetic parameters cannot be explained by age-related changes alone. Ince et al.  studied A new dosing schedule was designed, aiming for midazolam concentrations between 45 and 64 ng/mL for postoperative monitoring after procedural sedation  and betwExtracorporeal membrane oxygenation (ECMO) is a form of prolonged cardiopulmonary bypass used to support patients with life-threatening respiratory or cardiac failure . In neonECMO yields variability and lack of predictability in drug behaviour. The most common mechanisms by which ECMO affects pharmacokinetics are sequestration of the drug in the circuit and the increase of drug Cl and Vd , 56. TheAdequate sedation of neonates receiving ECMO is essential to allay the physical, emotional, and psychologic distress experienced during intensive care. Midazolam, a short-acting benzodiazepine derivative with rapid onset of action, is the sedative of choice and is preferred over other benzodiazepines because of its water solubility and the perceived rapid elimination .n = 10) received midazolam extracorporeally (into the circuit), via a pigtail catheter, whereas group 2 (n = 10) received the drug via a central or peripheral venous catheter. Midazolam was administered as a continuous infusion at a rate between 50 and 250 μg/kg per hour, initiated as soon as cannulation was achieved and extracorporeal blood flow was established. The pharmacokinetic parameters obtained by Mulla et al. [Twenty neonates with gestational age of 39.5 ± 1.9 weeks, postnatal age of 3.8 days, and body weight of 3,400 ± 600 g were enrolled in the study . The infa et al.  are summThe mean metabolic ratio was of 1-hydroxymidazolam to midazolam 0.17 (range: 0.03 to 0.9) and appeared to be significantly higher than that previously reported in non-ECMO infants (0.09), and it was similar to those observed in older children and adults .DeBerry et al.  determinμg/kg) was administered by a venoarterial catheter as a bolus infusion before cannulation. The median ECMO duration was 124 hours (range: 70 to 275 hours). When discomfort occurred, midazolam was started as a continuous infusion of 100 μg/kg per hour. The midazolam dose was incrementally adjusted (with steps of 100 μg/kg per hour) on the basis of the required level of sedation, which was objectified with the validated COMFORT behaviour scale. Once the patient was fully sedated, the midazolam infusion was interrupted until COMFORT scores indicated that additional sedation was necessary.Ahsman et al.  studied Midazolam Cl and Vd rapidly increased in 5 days of ECMO therapy. The kinetic parameters of midazolam are summarised in μg/kg per hour for the first 6 hours, after which, the infusion rate should be reduced to 150 μg/kg per hour. After 5 days, the infusion rate should be increased to 200 μg/kg per hour to compensate for the continual increasing of midazolam Cl and 1-hydroxymidazolam Cl. 1-Hydroxymidazolam glucuronide accumulated during ECMO, providing an increased proportion of the overall effect, up to 34% after 7 days of ECMO therapy. Large unexplained interindividual variability warrants careful titration of sedation effects. Ahsman et al. [To quickly reach stable serum concentrations of 400 ng/mL, the optimal dose regimen would be a continuous infusion of 300 n et al.  did not n et al. , 61.μg/kg per hour (range: 11 to 603 μg/kg per hour) to 109 infants; the infusion lasted for 6 to 171 hours (62 ± 31 hours). A bolus dose of 210 ± 239 μg/kg  was administered to 22 infants, and a combination of both was administered to 56 neonates. The mean midazolam doses required for critically ill neonates are lower than those required for wealthy neonates. Mean t1/2 was 9.9 hours, Cl was 1.2 ± 0.2 mL/min/kg, and Vd was 1.0 ± 0.2 L/kg [r = 0.89; P = 0.0001). The 531 measured plasma concentrations of midazolam ranged from 0 to 7,100 ng/mL; 105 samples were above 1,000 ng/mL. Midazolam was not detected in 19 samples. Aminoglycosides, aminopenicillins, and third generation cephalosporins increased the plasma midazolam concentrations for 73.8%, 72.2%, and 48.7%, respectively. Aminoglycoside treatment was associated with a 50% decrease in midazolam Cl.In literature there are three studies of midazolam population pharmacokinetics –34. One μg/kg was administered every 4 to 6 hours. Average parameter values for infants with birthweight of 1,000 g were as follows. Systemic Cl was 0.783 mL/min (CV = 83%) and Vd of central comportment was 473 mL (CV = 70%). t1/2 was not given. For infants with body weight more than 1,000 g, Cl was 1.24 mL/min (CV = 78%) and Vd was 823 mL (CV = 43%). The average population Cl in boys (0.642 mL/min) was less than in girls (0.808 mL/min). The level of significance of this difference was not given. The population mean Cl of overall patients was 0.938 mL/kg per min. The average Vd of overall infants was 1.15 L/kg, which agreed closely with the Vd of 1.0 L/kg in term and preterm infants [Lee et al.  studied  infants .μg/mL followed by a continuous infusion ranging from 50 to 400 μg/kg per hour, for 3.8 hours to 25 days for conscious sedation. t1/2, Cl, and Vd of midazolam were 5.5 ± 3.5 hours, 5.0 ± 3.9 mL/kg per min, and 1.7 ± 1.1 L/kg, respectively. Half of the patients had an age less than 6 months [The population kinetics of midazolam was studied in eighteen pediatric patients, aged from 2 days to 17 years . Midazol6 months . Mean Cl6 months , 62).The Cl reported by de Wildt et al.  is compa1-Hydroxymidazolam to midazolam ratio  is in aVery little is known about the pharmacodynamics of midazolam in infants and only two articles are available in literature , 63. Swaμg/kg per hour, for a period of 3.8 hours to 25 days [μg/kg per hour) were in agreement with other data from studies in pediatric intensive care patients [Twenty-one infants, aged from 2 days to 17 years, received a continuous infusion of midazolam from 50 to 400  25 days . A possi 25 days  recommen 25 days . As reflpatients , 71.Neonates can perceive pain when mechanically ventilated: this fact has been demonstrated by the physiological, biochemical, and behavioural changes indicative of pain and stress in newborn infants during mechanical ventilation . Pain inD. A. Rosen and K. R. Rosen  wrote a The American Academy of Pediatrics Committee on Drugs has issued guidelines categorizing pharmacological intervention into three levels: conscious sedation, deep sedation, and general anaesthesia . ConscioIntravenous infusion of benzodiazepines is important to assist ventilation and other potentially distressing procedures. Combined with intermittent or continuous infusion with opioids, the benzodiazepines provide smooth control of anxiety, pain, and agitation . The majμg/kg followed by a maintenance dose ranging from 37.5 to 100 μg/kg per hour over 47 hours. The sedation procedure was classified as a success if all the following clinical criteria were met: no agitation, no grimacing, and no crying facial expression before as well as during tracheal suctioning. The estimated success probability, updated after each patient, was <95% for all dose levels and led to allocation of the maximum loading dose (200 μg/kg). Based on the 23 patients studied, the final estimated probability of success was 76.9%  for the 200 μg/kg loading dose. The posterior success probability for the six doses 75, 100, 125, 150, 175, and 200 μg/kg corresponded to 27.7%, 32.7%, 38.1%, 47.3%, 52.5%, and 76.9%, respectively. Failures were recorded in 5 (25%) of 20 patients receiving a 200 μg/kg midazolam loading dose. No significant effect of sedation on pulse oximetry, oxygenation index, or triggering of ventilator breathing was evident after the onset of midazolam administration. No significant adverse effects were observed. Pneumothorax was reported in two children who had received a 200 μg/kg initial loading dose and were classified as “success” for sedation. A decrease of more than 30% for systolic, diastolic, and mean blood pressure was observed during the study inclusion in none, two (8.7%), and one (4.3%) children, respectively .Treluyer et al.  determinn = 24) or placebo (n = 22) as a continuous infusion [μg/kg per hour of midazolam. Infants below 33-week gestation received the same continuous infusion of 60 μg/kg per hour during the first 24 hours of treatment, followed by a continuous infusion of 30 μg/kg per hour of midazolam. The assigned treatment lasted for 5 days, but sedation with midazolam could be continued if prescribed by the attending neurologist. Midazolam gave a significantly better sedative effect than placebo (P < 0.05). Heart rate and blood pressure were reduced by treatment but remained within the normal range for gestational age and there was no effect on ventilatory indices. Hemodynamic instability requiring albumin and/or vasoactive drugs occurred in 14 infants, 8 of whom were in the midazolam group and 6 infants were in the placebo group. Intraventricular haemorrhage occurred in 8 infants in the midazolam group and 6 infants in the placebo group. Blood samples were obtained from 23 of 24 infants. Mean midazolam concentrations were 634 ± 234 ng/mL, 628 ± 327 ng/mL, and 543 ± 327 ng/mL at 24 and 48 hours and at the end of treatment, respectively. Substantial variability in plasma concentrations of midazolam was seen, but none of the infants had values below 200 ng/mL and only 2 infants  had a plasma concentration of midazolam above 1000 ng/mL. There was no correlation between sedation score and corresponding midazolam concentration. No midazolam-related adverse effects were noted. Continuous infusion of midazolam at doses adapted to gestational age induces effective sedation in newborn infants on mechanical ventilation, with positive effects on hemodynamic variables. Midazolam was given over only a few days and the limited effects on heart rate and blood pressure that were reported should not encourage long-term administration.Forty-six infants undergoing mechanical ventilation for respiratory distress syndrome were randomly assigned to receive midazolam  for analgesia and sedation [Midazolam was administered as a continuous infusion at a dosage of 50 to 400 sedation . The meaμg/kg followed by continuous infusion of 60 μg/kg per hour. Sixteen infants received placebo. The midazolam group had significantly better sedation than the placebo group from 18 to 24 hours after enrollment. At 48 hours, there were no significant differences in sedation between midazolam and placebo groups. None of the infants had hypotension on loading midazolam. Sedation provided by continuous infusion of midazolam appeared to be comparable to morphine alone in newborn infants on mechanical ventilation. These authors observed that the behavioural assessment was used to demonstrate the sedative effect of midazolam in ventilated newborn infants. Opioid and benzodiazepines combination is most commonly used to sedate newborns, but their use is limited by prolonged variable elimination of these drugs, leading to prolonged sedation. Midazolam produces a rapid and consistent effect because of its short t1/2. Hypotension has been reported as an adverse effect of midazolam in few infants after intravenous administration of midazolam [Arya and Ramji  performeidazolam , 78. Aryidazolam  did not When minimal sedation is required, intranasal midazolam is an excellent sedative . IntranaIntranasal midazolam use has been reported since 1988 . The intμg/kg (range: 300 to 800 μg/kg). Laceration repair was the most common procedure necessitating sedation (89%). Eleven patients (5.4%) required an additional sedative dose to complete the procedure. Ten of 11 patients received ketamine as the adjunctive sedative, and 1 patient required additional intranasal midazolam. There was 1 adverse event (0.5%). This was a minor desaturation episode following ketamine administration requiring a brief blow by oxygen. There were no adverse events in patients who received intranasal midazolam. These authors conclude that atomized intranasal midazolam is effective in providing anxiolysis to children undergoing minor procedures in the pediatric Department.Intranasal midazolam was studied by Lane and Schunk  using a μg/kg. Intranasal midazolam has a rapid onset and is more convenient than intravenous agents, because of its short duration. A dose of 200 μg/kg is safe and effective. In 142 of 162 cases (88%), midazolam demonstrated a positive effect; in 20 cases (12%), it was ineffective. The adverse effects were brief coughing, sneezing, or crying often occurring immediately after midazolam administration, but these were not considered adverse reactions unless they persisted. In 91 of 162 cases (56%), midazolam was used in conjunction with another agent. Fifteen adverse reactions (9%) occurred, including delay in awakening in 4 patients, reactive airway in 2 cases, desaturation and apnea in 4 cases, seizure in 1 case, agitation in 2 cases, and vomiting in 2 cases. Midazolam was the sole sedative in 71 of 162 (44%) cases. The use of midazolam to minimize the difficulty of starting an intravenous infusion constitutes the most common application. In 89% of the patients, it accomplished the desired effect. Of particular interest was the use of midazolam as a primary means of sedation since its intranasal route obviates the need for intravenous medication. An intranasally administered sedative with quick onset and short duration seems very innocuous.Harcke et al.  reviewedμg/kg was administered to 15 preterm infants with gestational and postnatal ages ranging from 26 to 31 weeks and from 3 to 13 days, respectively. Mean ± SD of body weight was 1076 ± 240 g. In 8 out of these 15 patients, the pharmacokinetics of intravenous midazolam was also studied. Apparent oral Cl, apparent Vd, and plasma t1/2 are given as median and range. Cl was 2.7 mL/kg/min (range: 0.64 to 15.5 mL/kg/min), Vd was 1.4 L/kg (range: 0.3 to 12.1 L/kg), and t1/2 was 7.6 hours (range: 1.2 to 15.1) hours. The absolute bioavailability was 0.49 (range: 0.12 to 1.0).Data describing midazolam disposition following oral administration to preterm infants are few. In literature, there are only two articles on the fate of midazolam following oral administration to neonates , 6. A siμg/kg midazolam. Pharmacodynamic measurements consisted of a COMFORT score at baseline and at 0.5, 1, 2, 4 and 6 hours after dose. A total of 24 infants were enrolled, of whom seven received both intravenous and oral midazolam, 13 infants received only intravenous midazolam, and 4 received only oral midazolam.In another study, De Wildt et al.  evaluateP = 0.003). No relationship was found between overall COMFORT scores or change in COMFORT score from baseline and midazolam. Diastolic blood pressure decreased significantly (approximately 11%) after intravenous, but not after oral, midazolam administration. No serious adverse events were reported. Midazolam administered as an intravenous infusion or oral bolus dose appears to be effective and well tolerated in a small majority of preterm infants. However, a considerable number of neonates did not respond to midazolam. The lack of response may be due to the fact that patients truly experienced therapeutic failure and/or consequent to the inability of the COMFORT score to adequately reflect sedation uniformly in sick preterm infants.Overall, mean COMFORT score decreased  significantly within 30 min after intravenous administration and within 1 hour after oral administration . If seizures continue, consider phenobarbital drip or lidocaine drip if not yet tried (unless phenytoin/fosphenytoin has been used).Slaughter et al.  reviewedThe neonatal period is one of the highest risk periods for seizures , which oμg/kg per hour for a period of 43 to 54 hours. Midazolam may be considered a safe and effective antiepileptic drug in refractory neonatal seizures of diverse aetiologies.Van Roij et al.  reviewedStatus epilepticus, a serious, life-threatening emergency, characterized by prolonged seizure activity, occurs most commonly in pediatric patients. Diazepam, phenytoin, or phenobarbital terminates seizures within 30 to 60 min, but there are patients who are refractory to standard therapy. Midazolam has emerged as a new treatment option. The review by Holmes and Riviello  comparedμg/kg per min, increasing by 0.5 to 1 μg/kg per min every 2 min until a favourable response or a maximum of 18 μg/kg per min). If the status epilepticus persisted after the initial bolus, another bolus of 100 to 150 μg/kg was administered 15 to 30 min later. Nonresponders to phenobarbital/phenytoin were controlled by midazolam.Castro Conde et al.  stated tLevel 1: rectal midazolam of 500 μg/kg, or 100 μg/kg, intravenously was administered. Level 2: after 10 min, 20 mg/kg phenytoin was administered intravenously. Level 3: after phenytoin load, 200 μg/kg midazolam was administered intravenously as a bolus, followed by a continuous infusion of 100 μg/kg per hour of midazolam, increased by 100 μg/kg per hour every 10 min, after extra loading of 100 μg/kg, to a maximum of 1 mg/kg per hour. Level 4: phenobarbital 20 mg/kg intravenously, or pentobarbital 2 to 5 mg/kg intravenous load, 1 to 2 mg/kg per hour of phenobarbital was continuously intravenously infused. Eighty-two of 122 infants (68%) received initial rectal diazepam. Cessation of epileptic activity was achieved with midazolam in 58 patients (48%). Midazolam is an effective agent for seizure control [The efficacy of a combination of midazolam and phenytoin in treating generalized convulsive status epilepticus was studied in 122 children with the median age of 24.4 months (range: 0.5 to 197 months) . Forty-t control . Several control , 102–104μg/kg per min, with an average of 4 μg/kg per min. There were no significant changes in serum sodium, potassium, calcium, or glucose in any of the patients. Four patients had recurrent seizures, which stopped after midazolam was reinstituted. Adverse effects included hypotension in 12 patients (38%), that was successfully controlled with dopamine and/or dobutamine, and transient urinary retention in 38% of infants. All the patients enrolled in the study had a successful management of seizures. These results suggest that midazolam is safe and effective for the treatment of uncontrollable seizures.Hu et al.  determinMidazolam is a highly effective antiepileptic agent, even in patients who have not responded to other benzodiazepines . Compareμg/kg and then maintained between 100 and 400 μg/kg per hour. After the loading dose, clinical seizures were no longer observed in four infants. The remaining became seizure free within one hour after the administration of midazolam. These results show that midazolam administrated by continuous intravenous infusion may be a valuable adjunct in the management of refractory neonatal seizures.Sheth et al.  studied McIntyre et al.  comparedMahmoudian and Zadeh  determinMany agents can depress the function of the central nervous system producing calming or drowsiness (sedation). Sedation may be performed with opioids , with the sedative-hypnotic midazolam and with the aesthetic propofol in ventilated neonates.Endotracheal intubation of premature infants is performed frequently in the “neonatal intensive care units” and delivery room. The procedure is associated with physiological and biochemical responses, and now there is strong evidence that premedication  improves the physiological stability, decreases the difficulty of procedure, and also reduces the potential for airway injury .μg/kg and 1 μg/kg remifentanil. The remaining 10 patients, with a gestational age of 31.5 ± 1.5 weeks and a body weight of 1523 ± 35 g, were treated with 2,000 μg/kg propofol and 1 μg/kg remifentanil. All neonates were in nasal continuous airway pressure therapy before intubation, with similar ventilatory parameters and degree of respiratory system. No statistical differences were observed as well between the drugs regarding pain and stress level before and after the intubation using the NIPS scale. No differences were observed in heart rate and blood pressure between the two drugs. Adverse effects such as hypotension were observed in two infants in each drug, and bradycardia was seen in one infant in the propofol group. The present findings did not show any difference in the quality of intubation, presence of adverse effects, changes on the hemodynamic variables, and quality of sedation and analgesia achieved when midazolam or propofol was used as a hypnotic associated with remifentanil as premedication for tracheal intubation in preterm neonates. The pharmacokinetics of propofol was studied in 25 neonates after a bolus injection of 3000 μg/kg [Penido et al.  compared00 μg/kg  and founμg/kg) and midazolam (500 μg/kg) 30 min before induction of anaesthesia. Anaesthesia was induced with sevoflurane (1 μg/kg), followed by a bolus of sufentanil (1 μg/kg) and pancuronium (200 μg/kg). The groups were comparable with respect to sex, age, weight, type of surgery, cardiopulmonary bypass, aortic cross clamp, and circulatory arrest time. The values for acyanotic patients were similar with midazolam, propofol, and sevoflurane. The corresponding values for cyanotic patients were similar to the three anaesthetics. There was a significant correlation between ventilator hours and troponin T concentrations, at the time of 8 hours, in the sevoflurane group (r = 0.45) and midazolam group (r = 0.50) but not in the propofol group. The present findings show that the postoperative production of troponin T, in pediatric patients undergoing cardiac surgery, is similar to midazolam, propofol, or sevoflurane anaesthesia.Malagon et al.  comparedμg/kg per hour. Fifteen of the 24 children received a combination of midazolam with fentanyl; the latter was infused at a rate of 0.5 to 2 μg/kg per hour. The mean duration of fentanyl infusion was 64 hours (range: 20 to 188 hours). No cardiovascular or respiratory adverse effects were seen. After stopping the fentanyl infusion, the children were sedated with midazolam. With midazolam as the only sedative drug, a significantly (P < 0.05) higher mean infusion rate of midazolam was necessary to reach a reliable sedation and it was of 209 μg/kg per hour (range: 100 to 500 μg/kg per hour). Mean serum concentration of midazolam decreased from 513 μg/L  to 330 μg/L  after 4 to 5 days. Midazolam Cl increased in relation to the duration of midazolam therapy . In the early phase of midazolam therapy, Cl of midazolam was 5.8 + 3.8 mL/kg per min and increased significantly by 133% to 13.6 + 10.6 mL/min per min in the late phase of therapy. The reason for the increase of midazolam Cl may be either an increase of the Vd or an increase of the hepatic Cl due to an enzyme induction of midazolam or by increasing hepatic blood flow during a treatment over a longer period. During the first 24 to 72 hours of midazolam therapy, the sedation score was significantly correlated with the serum midazolam concentration . The desirable sedation score could be achieved during this time period at a serum concentration between 100 and 500 ng/mL with an infusion rate of 100 to 400 μg/kg per hour. Midazolam seems to guarantee reliable sedation and anxiolytic action. The evaluation of the sedation score showed that reliable sedation can be achieved at a midazolam infusion of 100 to 400 μg/kg per hour and at a serum midazolam concentration between 100 and 500 ng/mL. In combination with fentanyl, the midazolam infusion rate could be held at a lower level. Continuous intravenous infusion of fentanyl and midazolam has been shown to provide satisfactory analgesia.Hartwig et al.  studied n = 79; 1.05 ± 0.88 mg/kg per hour), midazolam , or both . In 18.5% of patients there were complex cardiac lesions. Pre- and postprocedure systolic and diastolic mean blood pressure were 72 ± 14 and 68 ± 12 mmHg, respectively. Mean procedure time was 79 ± 36.2 min. The anaesthesiologist's assistance was required by the cardiologist in 21 procedures. Pre- and postprocedure O2 saturation were 93.19 ± 8.72 and 93.63 ± 8.3, respectively. The mortality was zero. The two groups were not different in relation to the drug use (P = 0.283) or the complexity of the cardiac lesion (P = 0.051). However, there was significant difference between the two regarding the need for supporting drugs  or oxygen treatment . These authors conclude that low dose of ketamine and midazolam can be administered safely to most pediatric patients by the cardiologist, who can safely predict the need for an anaesthesiologist.Ketamine and midazolam are commonly used in children undergoing cardiac catheterization. Jobeir et al.  reviewedMidazolam is incompatible with fat emulsion, albumin, ampicillin, bumetanide, cefepime, ceftazidime, dexamethasone, fosphenytoin, furosemide, hydrocortisone succinate, micafungin, nafcillin, dimenhydrinate, perfenazine, ranitidine hydrochloride, thiopental, prochlorperazine edisylate, and sodium bicarbonate , 111. KeThere have been reports of life-threatening adverse respiratory and cardiovascular events occurring after administration of midazolam. Pain, tenderness, and thrombophlebitis have occurred following injection of midazolam. Hiccups have been reported. Death due to respiratory depression, hypotension, or cardiac arrest has been reported in patients given intravenous midazolam for conscious sedation .A receptors in the neonate [Midazolam is not free from adverse effects when administered to neonates. The first intravenous loading dose of midazolam administered to premature infants not infrequently causes respiratory depression, with hypotension, a fall in cerebral blood flow, and paradoxical agitation . The par neonate .Respiratory depression and hypotension are common when midazolam is used in association with narcotics or following rapid bolus administration. Hypotension has also been reported after continuous infusion of midazolam. The percentage of infants who develop hypotension varies with the studies. Seizure-like myoclonus has been reported in 8% of premature infants receiving continuous infusion . This alμg/kg midazolam, but the most disturbing observation was the appearance of involuntary epileptiform movements lasting for 15 to 30 sec. The pathogenesis of this phenomenon is speculative, but perhaps in these preterm infants the decrease in arterial pressure and heart rate has also an impact on cerebral blood flow or there is a direct effect of midazolam on the brain.Van den Anker and Sauer  observedNg et al.  recentlyThe major adverse effects associated with continuous infusion of midazolam are related to tolerance and an associated abstinence or withdrawal syndrome , 114. Wiμg/kg midazolam. No change in heart rate occurred during the study period, while mean arterial blood pressure decreased by 3 mmHg, 5 min after midazolam administration, compared to baseline values. A nonspecific fall in transcutaneous PCO2 was seen at 20 min after midazolam administration. Mean cerebral blood flow decreased from baseline by 12% at 5 min after midazolam administration then returned to predose values. Midazolam concentrations in the therapeutic range were proved to be effective in sedation of pediatric intensive care infants. As only minor cerebral and hemodynamic effects were found with the use of midazolam in stable ventilated preterm infants, it appears that midazolam is a safe, short-term sedative agent.Harte et al.  evaluateμg/kg midazolam, followed by a constant infusion of 200 μg/kg per hour, to 11 ventilated premature infants, caused changes in cerebral oxygenation and hemodynamics [μg/kg midazolam, followed by a continuous infusion of 150 μg/kg per hour. Harte et al. [μg/kg of midazolam to 10 infants with a birthweight ≤1500 g.The administration of a loading dose of 200 dynamics . Van Leudynamics  reportede et al.  reportedt1/2 of midazolam is 22-fold shorter than that of diazepam [t1/2, Cl, and Vd are 1.9 hours, 6.6 mL/kg per min, and 1.1 L/kg, respectively. In healthy preterm infants, t1/2 is 3.3-fold longer, Cl is 4.4-fold smaller, and Vd is 1.1 L/kg in adult and neonates. The rate of midazolam elimination is slower in infants than in adults. Midazolam is mainly eliminated by metabolism in adults. It is hydroxylated in position 1 to form 1-hydroxymidazolam by CYP3A4 and CYP3A5. These enzymes surge in the liver during the first weeks of life [t1/2 and the slower Cl observed in neonates than in older infants and adults.The present paper reviews the clinical pharmacology of midazolam; most of the work was performed in neonates, and several data have been obtained in children. Benzodiazepines provide effective control of patient agitation without imposing a significant load on the cardiovascular system. Midazolam is a short-acting benzodiazepine with rapid onset of sedation and is preferable to diazepam. diazepam . In adul of life , 9, 11 aChildren, with an age greater than 2 years, had a Cl of midazolam that ranged from 5.8 to 13.6 mL/kg per min (mean ± SD = 9.6 ± 3.5 mL/kg/min) , 47. Theμmol/L and treated with midazolam, had a t1/2, Vd, and Cl of this drug 6.3-fold longer, 14.5-fold greater, and 2.6-fold larger, respectively, [Disease affects the pharmacokinetics of midazolam. A patient suffering from renal failure, with malignant hypertension, and having a creatinine concentration of 691 ctively,  than thectively, . Multiplctively, . The sevμg/kg per hour [t1/2 1.9-fold longer than the normal value [t1/2. The long t1/2 of midazolam observed by Jacqz-Aigrain et al. [t1/2, Cl, and Vd of midazolam [Fifteen preterm infants, requiring mechanical ventilation, were treated with midazolam continuous infusion of 60 per hour . They haal value . Some ofal value . Fentanyn et al.  may reflidazolam , 119.t1/2 was 1.6-fold greater [t1/2 in neonates. The infants studied by Lee et al. [t1/2 observed by Lee et al. [t1/2, Cl, and Vd were 5.5 ± 3.5 hours, 5.0 ± 3.9 mL/kg per min, and 1.7 ± 1.1 L/kg, respectively. These parameters were 0.87-fold lower, 3.0-fold higher, and 1.5-fold larger than the normal values. These data suggest that the Cl of midazolam increases with the neonatal development.In literature, there are three studies on the population pharmacokinetics of midazolam –34. Burt greater  than the greater  and 2.2- greater  than nor greater . These fe et al.  had a sme et al. . The lone et al.  might, aThe data on midazolam pharmacodynamics in neonates are few and do not allow any conclusion. De Wildt et al.  did not Neonates may undergo uncomfortable procedures and may experience moderate to severe pain in the “neonatal intensive care unit.” The availability of an appropriate sedative that reduces the stress and pain is desirable. Midazolam is an effective sedative and represents a convenient choice among the sedatives because of its fast onset of action and the rapid termination of effect. The use of midazolam for newborn sedation remains empirical, and further research on the effectiveness and safety of midazolam in ventilated neonates is needed before its employment can be considered in routine clinical use , 111.μg/kg (range: 300 to 800 μg/kg). The bioavailability of intranasal midazolam ranged from 50% to 83% [Sedation of neonates with midazolam may be performed by intravenous administration (bolus or continuous infusion) or by intranasal administration when minimal sedation is required . The med% to 83% . A secon% to 83% , 6. No iμg/kg midazolam, the final estimated probability of success is 76.9%. Using lower midazolam concentrations, the probability of success decreased.Treluyer et al.  determinμg/kg per hour. Infants, below 33 weeks of gestation, received the same continuous infusion rate during the first 24 hours of treatment, followed by a continuous infusion of 30 μg/kg per hour of midazolam. Mean midazolam concentrations were 634 ± 234 ng/mL, 628 ± 327 ng/mL, and 543 ± 327 ng/mL at 24 and 48 hours and at the end of treatment. Remarkable variability in midazolam plasma concentrations was observed, but none of the infants had levels of midazolam below 200 ng/mL and only 2 infants had a plasma concentration of midazolam above 1000 ng/mL.Mulla et al.  administμg/kg per hour produced midazolam concentrations sufficient for sedation of infants [μg/kg, followed by a continuous infusion of 60 μg/kg per hour, to 17 infants with a body weight <2000 g, undergoing mechanical ventilation. No significant adverse effects were observed. The course of mechanical ventilation was not influenced by the use of midazolam. These authors observed that the behavioural assessment was used to demonstrate the sedative effect of midazolam in ventilated infants.A dose of midazolam between 50 and 400  infants . Dosage  infants  administμg/kg was administered to infants (n = 15) with gestational age ranging from 26 to 31 weeks. The apparent oral Cl and Vd and the plasma t1/2 (given as median) were 2.7 mL/kg per min, 1.4 L/kg, and t1/2 7.6 hours, respectively. These values are not considerably different from the normal values. The absolute bioavailability “F” of midazolam was 0.49.Data describing midazolam pharmacokinetics following oral administration to neonates are few; only two articles are available in literature , 6. A siThe neonatal period is one of the highest risk periods for seizures , which oThe efficacy of a combination of midazolam and phenytoin, in treating generalized convulsive status epilepticus, was studied in 122 children with the median age of 24.4 months . Forty-tμg/kg followed by a continuous infusion of 1 to 15 μg/kg per min. Twelve patients (38%) developed hypotension. Midazolam emerged as a treatment option against the status epilepticus in patients who did not respond to diazepam, phenytoin, or phenobarbital [Midazolam successfully managed seizures in 32 neonates who did not respond to diazepam, phenytoin, or phenobarbital . Midazolbarbital . Comparebarbital .Many drugs depress the function of the central nervous system producing sedation in ventilated infants. They are the opioids , the sedative-hypnotic midazolam, and the anaesthetic propofol. Mahmoudian and Zadeh  did not μg/kg midazolam followed by an infusion of this drug at a rate of 100 μg/kg per hour. Fifteen of the 24 infants received a combination of midazolam with fentanyl. The latter was infused at a rate of 0.5 to 2 μg/kg per hour. No cardiovascular or respiratory adverse effects were seen. Midazolam guarantees reliable sedation and anxiolytic action. With midazolam, as the only sedative drug, a significantly higher mean infusion rate of 209 μg/kg per hour (range: 100 to 500 μg/kg per hour) was necessary to reach a reliable sedation. In the early phase of midazolam therapy, Cl was 5.8 ± 3.8 mL/kg per min and increased by 133% to 13.6 ± 10.6 mL/kg per min in the late phase of therapy.Hartwig et al.  experienμg/kg per hour (n = 79) and midazolam infusion was 140 ± 90 μg/kg per hour (n = 35). There were no differences between the two groups regarding the need for supporting drugs or oxygen treatment. In 18.5% of patients there were complex cardiac lesions. No difference was found in relation to the drug used (P = 0.283) or the complexity of cardiac lesions (P = 0.051). These authors conclude that low dose of ketamine and midazolam can be administered safely to most pediatric patients by the cardiologist, who can safely predict the need for an anaesthesiologist.Ketamine was infused at a rate of 1050 ± 880 A receptors present in the neonate [Life-threatening adverse respiratory and cardiovascular events have been reported in adults after midazolam administration. Pain, tenderness, and thrombophlebitis have occurred following intravenous midazolam administration . In neon neonate .μg/kg midazolam reduces the mean arterial blood pressure by 12% at 5 min after midazolam [μg/kg per min, to treat seizures. Twenty-seven percent of the infants receiving a continuous midazolam infusion of 60 μg/kg per hour developed hypotension [μg/kg per hour [The major adverse effects associated with continuous infusion of midazolam are related to tolerance and an associated abstinence or withdrawal syndrome , 76, 112idazolam . Hypotenidazolam  observedotension . These iper hour .t1/2 of midazolam. It is likely that these effects are due to the lowering of CYP3A4 and CYP3A5 activities. We feel that further research is required to ensure that the doses of midazolam recommended for the treatment of neonates are evidence-based.In conclusion, midazolam is a safe and effective drug, which may be used as a sedative or as an antiepileptic for the treatment of infants who are refractory to standard antiepileptic therapy. Midazolam is particularly useful in the treatment of the status epilepticus in infants. Midazolam is also used as an anaesthetic, but information about midazolam as an anaesthetic is lacking in neonates. Midazolam is extensively hydroxylated by two cytochrome P-450 forms, namely, CYP3A4 and CYP3A5. These enzymes surge in the liver during the first weeks of life. Neonates have a lower expression of these enzymes than older infants and adults. Consequently, the metabolic rate of midazolam is lower in neonates. Diseases affect the pharmacokinetics of midazolam; they may reduce the Cl or prolong


Aspergillus awamori. Compared to the non-A. awamori-fermented LPP, the growth effects of A. awamori-fermented LPP on Lactobacillus bulgaricus and Streptococcus thermophilus were four and two times higher after 3 days of fermentation, respectively. Increased 1,1-diphenyl-2-picrylhydrazyl radical scavenging activity and DNA protection activity of litchi pericarp polysaccharides were also achieved after A. awamori fermentation. Moreover, the relative content of glucose and arabinose in LPP after fermentation decreased from 58.82% to 22.60% and from 18.82% to 10.09%, respectively, with a concomitant increase in the relative contents of galactose, rhamnose, xylose, and mannose. Furthermore, lower molecular weight polysaccharides were obtained after A. awamori fermentation. It can be concluded that A. awamori was effective in biotransforming LPP into a bioactive mixture with lower molecular weight polysaccharides and higher antioxidant activity and relative galactose content.This study was conducted to increase the bioactivity of litchi pericarp polysaccharides (LPPs) biotransformed by  Litchi chinensis Sonn.) is a tropical-to-subtropical fruit, with a large amount of polysaccharides in its pericarp, pulp, or seed. As a major byproduct of litchi fruit, the pericarp accounts for approximately 16% of the whole fresh fruit and contains a significant amount of polysaccharides. Due to the health benefits, plant polysaccharides have become a desirable supplement in the functional food industry [In vitro studies have indicated that polysaccharides possess broad biological activities including antitumor [Lentinus edodes, Tremella fuciformis, and Astragalus membranaceus could selectively enrich beneficial bacterial species such as lactobacilli and bifidobacteria in an in vivo study [Litchi  on Lactobacillus bulgaricus and Streptococcus thermophilus after A. awamori fermentation.Previous c effect –12 and ic effect , 8, but c effect . Howeverc effect  but it icosidase . Thus, aLitchi chinensis Sonn.) cv. Huaizhi were harvested from a commercial orchard in Guangzhou, China. The fruit were washed with distilled water and then peeled manually. The pericarp was collected, then dried outdoors, and finally ground into powder.Fresh fruit of litchi  at 30°C. A. awamori spores were collected by washing the agar surface with sterile distilled water containing 0.1% Tween 80. The spore suspension was adjusted to a concentration of ca. 106 cfu/mL using sterile distilled water and then used to biotransform LPP.The microorganism All chemicals to prepare the Czapek-Dox medium were obtained from Guangzhou Reagent Co. 1,1-Diphenyl-2-picrylhydrazyl (DPPH), xylose (Xyl), arabinose (Ara), glucose (Glc), galactose , fructose (Fru), mannose (Man), fructose (Fru), and rhamnose (Rha), and dextrans standards were purchased from Sigma Chemical Company .Polysaccharides were extracted from litchi pericarp according to the method of Kong et al.  with min3 (30 mg), K2HPO4 (10 mg), KCl (5 mg), MgSO4·7H2O (5 mg), FeSO4 (0.1 mg), sucrose (0.3 g), LPP (0.3 g), and distilled water (10 mL). The medium was sterilized for 30 min at 121°C and then incubated with l mL of fresh A. awamori spore suspension at 28°C. The fermented products were collected by adding 50 mL of distilled water and holding the mixture fro 4 h at 70°C prior to filtering. This extraction was repeated, and the filtrates were collected, combined, and then concentrated under vacuum at 65°C. A control sample was prepared by incubating the fresh spore suspension (1 mL) with the previous culture medium but without LPP.For the fermentation, polyurethane foam (3 g) was placed into Erlenmeyer flasks (250 mL) and then mixed with the modified Czapek-Dox medium (10 mL) containing NaNO2PO4 (pH 6.0) at a flow rate of 0.6 mL/min. The column temperature was maintained at 35°C, and the injection volume was 30 μL. Dextrans with different molecular weights  were employed as standards. The molecular weights of the fermented and nonfermented LPPs were obtained from the equation based on the elution volume of standard dextrans to their log molecular weights.The molecular weight (MW) of LPP was determined by gel permeation chromatography (GPC) by the method of Yang et al. . The anaμL) was analyzed by gas chromatography. The temperature program started at 130°C, held for 1 min, increased to 180°C at 2°C/min, held for 3 min, increased to 220°C at 10°C/min, and held for 3 min [A gas chromatograph  equipped with a RTX-5 capillary column and a flame ionization detector was employed for the identification of the monosaccharides in the fermented and nonfermented LPPs. The fermented and nonfermented LPPs (10 mg) were hydrolyzed for 6 h with 10 mL of 2 M trifluoroacetic acid (TFA) at 120°C . After hThe method described by Sánchez-Moreno et al. was used to assess the DPPH radical scavenging activity of the fermented and nonfermented LPPs . BrieflyEscherichia coli DH5a cells were transformed with pUC19 plasmid DNA and then grown overnight in the Luria-Bertani (LB) medium containing ampicillin (50 μg/mL) at 37°C. Plasmid DNA was purified using the UNIQ-10 Plasmid Kit . Evaluation of the protection activity against Fenton-reaction-mediated DNA breakage was conducted using supercoiled plasmid DNA [μL of 50 mM hydrogen peroxide, 10 μL of 0.2 μg/μL plasmid DNA, and 1 μL of samples at 0.05, 0.1, 0.2, 0.5, and 1 mg/mL were mixed. The reaction was initiated by adding 2 μL of 5 mM ferrous sulfate into the mixture. After 15 min at 30°C, the reaction was stopped by adding 10 μL of stop solution containing 8 M urea, 50% sucrose, 50 mM EDTA, and 0.1% bromophenol blue and then separated by 1% agarose gel electrophoresis for 40 min under 120 V condition. The agarose gel was stained with 0.05% (w/v) ethidium bromide and then analyzed with an image analyzer .The protective abilities of the fermented and nonfermented LPP on supercoiled DNA damage were investigated by the method of Kang et al. . Escherismid DNA . Five miL. bulgaricus and S. thermophilus were conducted by the method of Yang et al. [μL) of L. bulgaricus or S. thermophilus cells in 10% (v/v) glycerol/water solution was pipetted to Man, Rogosa, and Sharp (MRS) agar plate containing 10 g/L protease peptone, 10 g/L beef extract, 5 g/L yeast extract, 1 g/L Tween 80, 2 g/L ammonium citrate, 5 g/L CH3COONa, 0.1 g/L MnSO4, 0.05 g/L MgSO4, 2 g/L K2HPO4, 20 g/L glucose, and 13 g/L agar, then inoculated at 37°C, and finally incubated for 48 h. A single colony was then transferred to a tube containing 5 mL of sterilized MRS broth  and then cultured for 24 h at 37°C by shaking at 130 rpm. To comparatively evaluate the growth effect, an aliquot (100 μL) of L. bulgaricus or S. thermophilus cell suspension (starting concentration of 1 × 106 cfu/mL) was mixed with 2.5 mL of MRS broth and LPP samples at 0, 50, 100, and 200 μg/mL. The mixture was incubated for 24 h at 37°C with shaking at 130 rpm. After dilution, the mixture (100 μL) was added to MRS agar plate and then cultured at 37°C for 48 h. The final cell concentration was estimated by counting the colonies on the plate.The growth effects of the fermented and nonfermented LPPs on g et al. . An aliqData were expressed as mean ± standard deviations (SD) and then analyzed by OriginPro 8 . Graphs were made using Sigmaplot . One-way analysis of variance (ANOVA) and the Tukey's multiple comparisons were carried out to test for significant differences between the means. Differences between the means at the 5% level were considered significant.V + 0.459 V 2 − 0.00938 V 3 and the correlation coefficient of 0.997 . As shown in A. awamori-fermented LPPs displayed two peaks in the chromatogram, and their molecular weights were estimated to be 98.524 and 24.441 kDa, respectively. After 3 days of fermentation, one band with an estimated MW of 69.736 kDa was observed , arabinose (18.78%), galactose (9.99%), rhamnose (5.03%), mannose (3.79%), and xylose (2.49%). The result was different from the previous report by Yang et al. [A. awamori can produce a number of arabinanases [The monosaccharide profiles of non-alactose .99%, rhainanases  and glucinanases  that mayinanases , which cA. awamori-fermented LPP, less than 10% of the DPPH radical scavenging activity was observed when a concentration of 1 mg/mL was used, which was consistent with the previous reports in the polysaccharides extracted from litchi seed and soy sauce lees, respectively [A. awamori-fermentation, the LPP exhibited a marked increase in the DPPH radical scavenging activity. After 3 and 6 days of fermentation, the A. awamori-fermented LPP had a stronger DPPH radical scavenging activity than the non-A. awamori-fermented LPP. When 1 mg/mL was employed, the DPPH radical scavenging activity of the A. awamori-fermented LPP increased from 9.24 ± 0.53% on the initial day to 17.47 ± 0.35% on the third day and to 18.27 ± 0.27% on the sixth day, which suggested that the polysaccharide modification influenced greatly the DPPH radical scavenging activity. Previous report of Kong et al. [Epidemiological evidence indicates an inverse correlation between the consumption of plant antioxidants and incidence of several chronic diseases . The DPPectively , 6. AfteA. awamori-fermented and non-A. awamori-fermented LPPs were used to further investigate the DNA protection activity. As shown in A. awamori-fermented LPP (Line B to F) and A. awamori-fermented LPP after 3 days (Line H to L) and 6 days (Line M to Q) exhibited an increased DNA protection activity against supercoiled breakage after fermentation, which further confirmed that the fermentation enhanced the antioxidant activity of LPP  the growth of L. bulgaricus. A dose-dependent response was observed in the present study. Compared to the non-A. awamori-fermented LPP, the fermented LPP at 50 μg/mL after 3 days of fermentation showed four times higher growth effect, but no significant difference was observed between the non-A. awamori-fermented and A. awamori-fermented LPPs at between 50 and 100 μg/mL after 6 days of fermentation. Similarly, the A. awamori-fermented LPP after 3 days of fermentation showed much higher growth effects on S. thermophilus compared with the non-A. awamori-fermented LPP (A. awamori-fermented LPP after 6 days of fermentation decreased. The present study demonstrated that the A. awamori-fermented LPP after 3 days of fermentation with an MW of 69.736 kDa possessed the higher growth effects on L. bulgaricus and S. thermophilus compared to the non-A. awamori-fermented LPP (about 98.524 kDa) and the fermented LPP (about 9.844 Da) after 6 days of fermentation. Previous studies have also indicated that low weight polysaccharides or hydrolyzed oligosaccharides can enhance these effects [The growth effect of non-nted LPP . However effects .Bifidobacteria, Lactobacilli, and Streptococcus densely populates in human gut. The microorganisms belonging to Lactobacilli genus are believed to be beneficial to human health through improving absorption of nutrients, preventing colonisation of pathogens and stimulating immune response in humans [Aspergillus awamori-fermented litchi pericarp polysaccharides on L. bulgaricus and S. thermophilus in an effort to understand the beneficial ability of the biomodified litchi polysaccharides. However, the relationship between biomodified litchi pericarp polysaccharide and growth effects on L. bulgaricus and S. thermophilus needs to be further investigated.Large amount of microorganisms including n humans . Thus, wA. awamori can degrade markedly polysaccharide with reduced molecular weight and increased contents of galactose, rhamnose, mannose, and xylose. Increases in growth effect of L. bulgaricus and S. thermophilus, DNA protection activity, and DPPH radical scavenging activity were obtained after fermentation of LPP by A. awamori. This study provided evidence that litchi paricarp polysaccharide can be enzymatically modified to increase its bioactivity. Thus, A. awamori fermentation of LPP can be served potentially as a means of increasing the utilization of this readily accessible waste material.The fermentation of LPP by


Aim of the study was to compare the effect of neuroleptic sedation with acepromazine and neuroleptanalgesia with acepromazine and buprenorphine on thermal thresholds (TT) obtained at the nostrils and at the withers. The study was carried out as a randomized, blinded, controlled trial with cross-over design. Thermal thresholds were determined by incremental contact heat applied to the skin above the nostril (N) or the withers (W). Eleven horses were treated with saline (S), acepromazine (0.05 mg/kg) (ACE) or acepromazine and buprenorphine (0.0075 mg/kg) (AB) intravenously (IV). Single stimulations were performed 15 minutes prior and 15, 45, 75, 105, 165, 225, 285, 405 and 525 minutes after treatment. Sedation score, gastrointestinal auscultation score and occurrence of skin lesions were recorded. Data were analysed with analysis of variance for repeated measurements.There were no significant differences in TT between N and W with all treatments. The TT remained constant after S and there was no difference in TT between S and ACE. After AB there was a significant increase above baseline in TT until 405 minutes after treatment. Restlessness occurred 30–90 minutes after AB in 7 horses. All horses had reduced to absent borborygmi after AB administration for 165 to 495 minutes.Thermal stimulation at both described body areas gives comparable results in the assessment of cutaneous anti-nociception in horses. There is no differential influence of neuroleptic sedation or neuroleptanalgesia on TTs obtained at N or W. Buprenorphine combined with acepromazine has a long lasting anti-nociceptive effect associated with the typical opioid induced side effects in horses. Thermal stimulation is a natural modality to induce nociception. In addition, it is able to stimulate C-fibre nociceptors and is therefore suitable to detect opioid induced antinociception ,2. In hoHowever, the type of nocifensive response and its detection varies between body regions and might therefore influence the determination of thermal thresholds. Transmission of a noxious stimulus at the body travels through the dorsal horn of the spinal cord and ascends to the brain via the spinothalamic tract . AssessmTherefore, the aim of the study was to compare the effect of neuroleptic sedation with acepromazine and neuroleptanalgesia with acepromazine and buprenorphine on TTs obtained at the nostrils and at the withers. The hypothesis was that neuroleptic sedation and neuroleptanalgesia would inhibit conscious reactions to a greater extent than reflex responses.There was no significant change in skin temperature at N over the course of the experiment in group S or ACE, but in group AB there was a statistically significant increase in skin temperature over the first measurements Table . Skin teThermal threshold temperatures at the nostril did not change over the observation period in group S and group ACE Figure A. In groThe most frequent reaction to thermal stimulation at N was head shaking (54.2%) followed by rubbing the face (44.4%). Stimulation at W was mostly answered with skin twitching (86.7%) rarely with shaking the whole body (12.8%) or turning the head towards the stimulus (0.5%).All horses became mildly to moderately sedated  15 miGastrointestinal sounds were reduced or absent from 15 to 75 minutes after AB administration and lasted for 180 to 510 minutes (297 ± 124 min). The horse not showing signs of sedation in group AB developed signs of colic one hour after the experiment was completed. An impaction of the large colon was diagnosed and treated medically without further complications. In group AB, excitatory phenomena like restlessness and box walking were observed in 10 of 11 horses starting with waning sedative effects of acepromazine  was 6.4 hours  resulted in very variable threshold temperatures and skin lesions in horses . Reducin1-adrenolytic activity, depresses the vasomotor center followed by hypotension, vasodilatation and increased digital blood flow /[Tc ‒ T0]). TT is the thermal threshold temperature, T0 is the skin temperature and Tc is the thermal cut-out temperature [TE  %   =  100 ×  with repeated measurements within subjects and post-hoc Tukey-Kramer test for multiple pair wise comparisons. Statistical significance was attributed when a 12 SWG, EquiCathTM Fastflow, Braun Vet Care GmbH, Tuttlingen, Germany.b Scandicain®, AstraZeneca GmbH, Wedel, Germany.c Dafilon® 1 metric, B. Braun, Aesculap AG, Tuttlingen, Germany.d Topcat Metrology Ltd., Little Downham, Ely, UK.e 0.9% natriumchlorid solution, B. Braun Melsungen AG, Melsungen, Germany.f Vetranquil® 1%, Albrecht GmbH, Aulendorf, Germany.g: Temgesic®, Essex Pharma GmbH, München, Germany.h Vet-Sept®, Albrecht GmbH, Aulendorf, Germany.i Vacuette®, Greiner Bio-One GmbH, Kremsmünster, Austria.j Eppendorf cuvettes, Eppendorf- Netheler- Hinz GmbH, Hamburg, Germany.k Agilent 6400 Sereis, Triple Quad LC/MS system, Santa Clara, CA.l LIPIDOMIX GmbH, Dr. rer. nat. M. Rothe, Berlin, Germany.m PK Solutions 2.0 TM, Summit Research Services, Pharmacokinetics and Metabolism Software, Montrose, USA.n SAS® version 9.2, SAS Institute, NC, USA.o GraphPad Prism® version 5.03, GraphPad Software, Inc., CA, USA.AB: Acepromazine and buprenorphine; ACE: Acepromazine; N: Nostril; S: Saline solution; TE %: Percentage of thermal excursion; TT: Thermal thresholds in °C; W: Withers; WTT2: Wireless thermal threshold testing device 2 (modified for horses).The authors declare that they have no competing interests.SBRK conceived the project. CP accomplished the practical part of the study and was supported from KH and SBRK. KR carried out the statistical analysis. SBRK, KH and CP participated in interpretation of the study results. CP drafted the paper which was later revised by all co-authors through substantial contributions to the content of the paper. All authors read and approved the final manuscript.Side effect after AB administration. The first horse seen in the video showed unconscious head nodding two minutes after AB administration for 3 minutes. One horse was deeply sedated (score: 8). Further horses were restless and showed box walking 3 – 4 hours after AB administration.Click here for file


Ulva prolifera and the native macroalga Gracilaria lichvoides, both of which were collected from the coastline of East China sea. In laboratory experiments, the presence of G. lichvoides at 1.25 g wet weight L−1 significantly inhibited growth and photosynthesis of U. prolifera at concentrations of 1.25, 2.50, and 3.75 g wet weight L−1 (p<0.05) in both semi-continuous co-culture assays and in co-culture assays without nutrient supplementation. In contrast, although U. prolifera had a density effect on G. lichvoides, the differences among treatments were not significant (p>0.05). Culture medium experiments further confirmed that some allelochemicals may be released by both of the tested macroalgae, and these could account for the observed physiological inhibition of growth and photosynthesis. Moreover, the native macroalgae G. lichvoides was a stronger competitor than the opportunistic species U. prolifera. Collectively, the results of the present study represent a significant advance in exploring ecological questions about the effects of green tide blooms on the macroalgal community.Allelopathy, one type of direct plant competition, can be a potent mechanism through which plant communities are structured. The aim of this study was to determine whether allelopathic interactions occur between the opportunistic green tide-forming species  Ulva. Green tides have occurred almost in many enclosed marine water bodies, including in Europe, Australia, New Zealand, Hong Kong, the Philippines, Indonesia, India, Egypt, China, South Africa and Central America Ulva prolifera caused the world's largest green tide 2The introduction and spread of exotic species into the marine environment is considered to be a major threat to marine ecosystems, with potentially dramatic effects on biological diversity, productivity, habitat structure, and fisheries U. prolifera have been well documented, the mechanism by which it invades a community and its impact on native communities has received little attention. The few studies conducted to date have shown that in addition to having negative effects on tourism, large algal mats can have deleterious ecological effects. These effects include uncoupling of the biogeochemical cycles in sediments from those in the water column While the occurrence and spread of Invasive macroalgae can impact native species through competition for different resources such as light, space, or nutrients Ulva fasciata, Ulva lactuca, Ulva pertusa, and Ulva linza on the harmful microalgae Prorocentrum micans, Prorocentrum donghaiense, Heterosigma akashiwo, Alexandrium tamarense, and Chaetoceros gracileP. micans and Skeletonema costatum or Karenia mikimotoiU. linza could release allelochemicals that could inhibit the growth of the red macroalga Gracilaria lemaneiformis (unpublished data).The bulk of research on allelopathy has focused on macroalga-microalga interactions, especially in the red tide-inhibition realm. There is now a considerable amount known about allelopathic effects of green macroalgal such as U. linza, U. prolifera is a dominant species responsible for forming green tides. However, Liu et al. (2010) et al. (2011) Ulva strain of the 2008 green algal bloom in the Yellow Sea was not detected in the coastal waters of Qingdao in the following winter. Compared to U. linza, much less is known about the allelopathic ecology of U. prolifera, and to date no studies have assessed what happens to the structure and biodiversity of a community when this opportunistic species invades it. In fact, few studies have addressed allelopathic interactions in the marine environment and the function of secondary metabolites as defenses against pathogens or other competing plants. Thus, in this study we examined the allelopathic interactions between the opportunistic species U. prolifera and the native macroalga Gracilaria lichvoides, both of which were collected from the coastline of East China Sea. We performed a series of laboratory experiments under controlled conditions in which we co-cultured the two species and also cultured them separately . Physiological parameters such as algal growth, algal photosynthesis, nutrient assimilation, and changes of pH in the culture medium were examined to assess potential allelopathic effects between the tested macroalgal species.Like U. prolifera and G. lichvoides were collected from the coastline of East China sea, in May 2011. In the laboratory, the intact samples were washed several times with sterile seawater, sterilized with 1% sodium hypochlorite for 2 min, and then rinsed with autoclaved seawater. Both U. prolifera and G. lichvoides were pre-cultured aseptically in f/2 medium in an incubator without N or P supplement for 48 h before running experiments. The temperature was maintained at 15°C. Illumination was provided by cool-white flouorescent lamps at 100 µmol photons m−2 s−1 and on a 12∶12 h light: dark cycle. All cultures were shaken manually twice at the same time every day. The pH and salinity of the seawater used for experiment were 8.0 and 30 ppt, respectively.Floating specimens of G. lichvoides and U. prolifera, in the batch culture experiment, G. lichvoides (1.25 g wet weight L−1) was co-cultured with three different density of U. prolifera . The experiments were conducted in 500 ml flasks containing 400 ml of culture medium with 882 µmol L−1 NaNO3 and 32 µmol L−1 KH2PO4 at 15°C and 100 µmol photons m−2 s−1. During the period of experiments, nutrients were not added into any flask to supply the decreasing of the nutrients. In addition, a serial of semi-continuous experiments were also conducted by regularly adding nutrients to 882 µmol L−1 NaNO3 and 32 µmol L−1 KH2PO4 every 24 h, while the culture conditions were the same as described above. G. lichvoides and U. prolifera were individually cultured (monocultured) as controls. All experiments in this study were conducted at in triplicate, and aseptic techniques were used in all experimental steps. Flasks were also monitored for pH levels during the experiments. Measurements were taken using a pH probe equipped with an electrode . These experiments lasted for 96 h. The growth of macroalgae U. prolifera and G. lichvoides was estimated by monitoring changes in algal wet weight at 0 h, 48 h, and 96 h.To determine the allelopathic interactions between the fresh thalli of G. lichvoides and U. prolifera in sterilized seawater at a concentration of 10 g wet weight L−1 for 48 h without nutrient enrichment. Thereafter, the macroalgal thalli were removed and the macroalga-free culture medium was filtered through 0.45 µm acetate cellulose filters and diluted 2 and 4 times with sterilized seawater. The three gradient concentrations of culture filtrate of G. lichvoides or U. prolifera were used for experiments to study the effects of culture filtrates on fresh algal thalli of U. prolifera or G. lichvoides at a concentration of 1.25 g wet weight L−1. The media containing culture filtrates were resupplied with nutrients every 24 h, and the pH was adjusted to 8.0 by 2 mol L−1 HCl every day. The culture system was kept at 15°C with a light intensity of 100 µmol photons m−2 s−1 and a 12∶12 h light: dark cycle. The experiments lasted for 96 h, and the growth of macroalgae U. prolifera and G. lichvoides was estimated by monitoring changes in algal wet weight at 0 h, 48 h, and 96 h.Macroalgal culture medium was prepared by separately culturing 3–N) and phosphas (PO4–P) were analyzed photometrically using an AutoAnalyzer . Nutrient uptake rates were calculated as: NUR = (0C−tC) V/DW/t, where NUR is the nutrient uptake rate (µmol of nutrient g FW wt−1 h−1); 0C and tC are the nutrient concentrations (µmol L−1) at the beginning and the end of the experiment, respectively; V is the volume of water (L); FW is the algal fresh weight (g), and t is the time interval (h).During the experiments, water samples (5 ml) in batch culture were collected daily, filtered immediately through acetate cellulose filters, and frozen in polyethylene flasks for storage until analysis. Concentrations of nitrate (NOY(II)] of fresh algal thalli was measured using the pulse–amplitude modulated method on a Dual-PAM-100  connected to a PC running WinControl software and calculated as follows: Y(II) = (Fm′−Ft)/Fm′. The real-time fluorescence yield Ft is obtained by averaging the fluorescence readings within 0.2 s and the maximum fluorescence yield (Fm′) was detected when the samples were illuminated by actinic light of 100 µmol photons m−2 s−1.Simultaneously, the effective PSII quantum yield [The significance of variance between treatments and the control or among treatments was tested using a one-way ANOVA or a multiple comparison test. All tests were run using the software SPSS 17.0. The significance level was set at 0.05 for all tests unless otherwise stated.U. prolifera had no apparent effects on the growth  or photosynthesis  of G. lichvoides compared to the control (Y(II) of the mono-cultured G. lichvoides increased by 17.0±7.1% and 90.6±2.5%, respectively. In the three co-culture systems containing 1.25, 2.50, and 3.75 g wet weight L−1 of U. prolifera, G. lichvoides biomass increased by 16.0±2.8%, 14.7±4.6%, and 12.0±2.1%, respectively, and Y(II) increased by 85.4±4.5%, 84.4±4.2%, and 82.5±2.0%, respectively. In contrast, the presence of G. lichvoides at a concentration of 1.25 g wet weight L−1 had a dramatic negative effect on growth of U. prolifera at concentrations of 1.25, 2.50, and 3.75 g wet weight L−1  . Moreover, although the Y(II) of U. prolifera increased by 5.4±1.6%, 6.7±0.6%, and 8.4±0.7%, the effects of G. lichvoides on photosynthesis of U. prolifera were not significant compared to the control treatment .In the batch co-culture experiments without nutrient supplement, different quantities of  control . After 9G. lichvoides also grew well both in monoculture and in co-cultures with U. prolifera  values in the co-culture treatments increased by 90.1±1.6%, 88.0±2.4%, and 85.4±3.4%, respectively, although all of these values were lower than that of the control, which increased by 150.2±55.2%. However, one-way ANOVA indicted that these differences were not significant . In contrast, G. lichvoides had density-dependent effects on growth and photosynthesis of U. prolifera. After incubation for 96 h, the biomass of U. prolifera declined significantly by 18.0±5.7%, 14.3±9.5%, and 8.3±11.8%, respectively, compared to the control, which increased by 36.7±6.7% . However, Y(II) of U. prolifera did not change significantly .rolifera  in semi-3–N concentration in the monoculture of U. prolifera decreased more quickly (from 882 to 325.0±53.1 µm L−1) than that in the monoculture of G. lichvoides (882 to 359.4±47.1 µm L−1), except for the first 12 h. The NO3–N concentration in the monoculture of U. prolifera and in the monoculture of G. lichvoides was significantly correlated with the concentration in the co-culture systems; this relationship illustrates that NO3–N in the co-culture assays was absorbed jointly by G. lichvoides and U. prolifera (G. lichvoides (4.5±0.4 µmol N g−1 FW h−1) in monoculture experiment was lower than that of U. prolifera (4.8±0.4 µmol N g−1 FW h−1), but the difference was not significant .rolifera . During 4–P in the monoculture of U. prolifera was significantly correlated with that in the co-culture of G. lichvoides with U. prolifera. Moreover, the PO4–P concentration in the co-culture system declined to much lower levels  compared with that in the G. lichvoides monoculture (24 µm L−1) after 96 h. These results indicate that the PO4–P was mainly absorbed by U. prolifera in all co-culture assays (G. lichvoides (0.08±0.01 µmol P g−1 FW h−1) in monoculture experiment was dramatically higher than that of U. prolifera (0.25±0.01 µmol P g−1 FW h−1) .The concentration of POe assays . MoreoveThe pH of the culture medium used in both the monoculture and co-culture systems initially was 8.0. Over time, the pH values in all treatments increased slightly (no more than 1.12) in both the semi-continuous assays and in the co-culture assays without nutrient supplementation . A conceU. prolifera or G. lichvoides was cultured with macroalgal culture filtrates of G. lichvoides or U. prolifera, respectively. The G. lichvoides culture filtrate dramatically inhibited growth  and photosynthesis  of U. prolifera in comparison to the control  values of U. prolifera increased by 3.8±4.6%, 3.1±2.7%, and 1.9±1.7%, the 2 and 1 times diluted culture filtrates of G. lichvoides had significant effects on photosynthesis of U. prolifera. Moreover, the U. prolifera culture filtrate also caused significant inhibition of growth  and photosynthesis  of G. lichvoides compared with those of the control. In the monoculture, the biomass and Y(II) value of G. lichvoides increased by 23.0±7.1% and 150.2±55.2%, respectively. When treated with 4, 2, and 1 times diluted culture filtrate of U. prolifera, the biomass and Y(II) of G. lichvoides increased by only 15.7±3.6%, 11.3±2.1%, and 8.1±2.3% and 38.5±7.4%, 16.9±5.1%, and 1.3±0.1%, respectively. control . After 9et al. (2011) K. mikimotoi was suppressed by Gracilaria verrucosa mainly through competition for nutrients, especially nitrogen. Previous studies have reported that the green macroalgae Ulva spp., which have a high surface area/volume ratio, exhibit high rates of nutrient uptake. But many of these algae are often limited in their ability to concentrate and store nitrogen internally, and are, therefore dependent on a constant high level of nitrogen in the medium Gracilaria spp., on the other hand, have opposite qualities in that they show a high capability of storing nitrogen (partly as phycoerythrin), and require only pulse fertilization U. prolifera concentrations  Gracilaria spp., when cultured in the presence of Ulva cf. lactuca, was not due to shading or nutrient depletion, but seemed to be caused by competition for inorganic carbon or some type of allelopathy. In the present study, the fresh algae was incubated in 500 ml flasks containing 400 ml of culture medium, and the space was big enough for the sample to growth. Meanwhile, the experiment was conducted in an illuminated incubator at 100 µmol photons m−2 s−1 and the algae can get the light from all directions. Additionally, all cultures were shaken manually twice every day and the samples could change their positions in the culture medium. Based on mentioned above, it makes light limit unlikely for growth.Light competition was another mechanism that account for the species interactions. Tait and Schiel (2011) G. lichvoides or U. prolifera. However, in the culture filtrate experiments  was found in both experiments (G. lichvoides filtrate added to U. prolifera culture or U. prolifera filtrate added to G. lichvoides culture)  . These ret al. (1998) G. lichvoides had strong allelopathic effects on the opportunistic species U. prolifera when the co-culture concentration of G. lichvoides was one-third times higher than that of U. prolifera. This may explain why the dominant U. prolifera strain of the bloom was absent in all the water-derived cultures during the sampling period Although green tides are widespread and invade many macroalgal ecosystems, they have been largely neglected in studies of the maintenance of biodiversity. The effects of green tide blooms are varied and have been summarized by Fletcher (1996) Undaria pinnatifida, whereas the presence of a stable native algal canopy inhibited invasion. On the west coast of Vancouver Island in Canada, White and Shurin (2011) Sargassum muticum on native macroalgal richness. In the present study, the highest concentration of U. prolifera in the co-culture system was 3.75 g wet weight L−1, and U. prolifera at this concentration had no significant effects on G. lichvoides (1.25 g wet weight L−1). However, the effect of higher co-culture concentrations of U. prolifera on G. lichvoides should be investigated in the future, as higher concentrations could significantly impact the native species.The effects of the introduction and spread of exotic species on community richness can be positive Ulva (formerly Enteromorpha) linza–procera–prolifera complex U. prolifera, the causative species of the world's largest green tide, is distributed widely in the intertidal zones of shores and estuaries around the world because of its tolerance of a wide range of salinity and water temperature, its high growth rate, and its extraordinary capabilities for propagation U. linza could restrict growth and photosynthesis of G lemaneiformis, even when the co-culture density of U. linza was equal to that of G. lemaneiformis (unpublished data). The present study represents a significant advance in exploring ecological questions about the effects of green tide blooms on the macroalgal community. If hybridization between U. linza and U. prolifera occurred, a more destructive species could have more serious ecological effects on the marine community.Previous studies have reported that the green tide-forming species in the Yellow Sea were


Okra has different uses as a food and a remedy in traditional medicine. Since it produces many seeds, distribution of the plant is also quite easy. Although seed oil yield is low (4.7%), since the linoleic acid composition of the seed oil is quiet high (67.5%), it can still be used as a source of (UNSAT) unsaturated fatty acids. In this study, samples of okra grown in four different locations were analyzed to measure fatty acid and amino acid compositions. The content of the lipid extraction ranged from 4.34% to 4.52% on a dry weight basis. Quantitatively, the main okra fatty acids were palmitic acid (29.18–43.26%), linoleic acid (32.22–43.07%), linolenic acid (6.79–12.34%), stearic acid (6.36–7.73%), oleic acid (4.31–6.98%), arachidic acid (ND–3.48%), margaric acid (1.44–2.16%), pentadecylic acid (0.63–0.92%), and myristic acid (0.21–0.49%). Aspartic acid, proline, and glutamic acids were the main amino acids in okra pods, while cysteine and tyrosine were the minor amino acids. Statistical methods revealed how the fatty acid and amino acid contents in okra may be affected by the sampling location. Abelmoschus esculentus) is widely distributed in tropical to subtropical regions in Africa, Asia, Southern Europe, Mediterranean countries, and America. Okra is mainly grown as a vegetable in the plains of Egypt. It grows well under warm climatic conditions (temperatures above 26°C).  Okra  near the desert, M pod (Mansoura) near the Nile River, K pod (Kafr El-Sheikh) near the Mediterranean sea, and D pod (Dakahlia) near a lake. The pods were sun-dried, and their contents were analyzed.  Lipids were extracted using the method of Soxhlet . All solFatty Acid Methyl Ester Preparation. Fatty acid methyl esters (FAMEs) were prepared according to [3 , and the reaction was allowed to proceed for 5 min. FAMEs were extracted twice with 10 mL of petroleum ether and 10 mL of water being added to the mixture.rding to . An aliqμm; Supelco, Inc., Bellefonte, PA, USA) was used. The column parameters were as follows: initial column temperature was held at 40°C for 5 min after injection, 20°C/min to 220°C, and finally the temperature was held at 220°C for 30 min. Helium was the carrier gas, with the column inlet pressure set at 17 psi. The detector temperature was 250°C. For identification purposes, these analyses were also performed with a gas chromatograph  coupled with a 5973 mass spectrometer detector (Agilent) by using the same column described previously. FAMEs were analyzed using gas-liquid chromatography  equipped with a flame ionization detector. An SP-2560 fused silica capillary column . Fatty acid contents were expressed as the proportion of each individual fatty acid to the total amount of all fatty acids present in the sample. FAMEs were identified by using standards  and comparing their mass-spectral data with the mass-spectral database in the Wiley 7.0 library . The conjugated linoleic acid peaks were identified by comparison with the retention times of the reference standard  for amino acid analysis. Sample aliquots containing around 8–12 mg of proteins were placed in a 20-mL cuvette and mixed with 9 mL of 6 M HCl . After sP < 0.05). The correlation between all the studied parameters was determined by the principal component analysis (PCA) using XLSTAT software.  Data from the replications of all varieties were subjected to a variance analysis (ANOVA) using SPSS 16.0 for Windows. Significant differences between the means were determined by Duncan's new multiple range test  was higher than that of the essential amino acids (E). M pod and K pod were found to be rich in isoleucine, lysine, and valine, with a combined concentration of 1.32 g/100 g for M pod and 1.46 g/100 g for K pod. However, significant differences (P < 0.05) in arginine, aspartic acid, and proline contents were observed between M pod and K pod. The amount of sulfur-containing amino acids (methionine and cystine) was 0.24, 0.23, 0.30, and 0.19 g/100 g for S, K, M, and D pod, respectively, while the total aromatic amino acid content was 0.66–0.96 g/100 g. D pod showed the lowest value, and M pod showed the highest value.  The amino acid profile of the okra plants is shown in  PCA was used to analyze the fatty acid and amino acid contents. Figures  The fatty acid and amino acid results for okra pods were in agreement with the literature. Quantitatively, total lipids ranged from 4.34 g/100 g to 4.52 g/100 g on a dry weight basis. Okra contents had a significant correlation with the geographical distances. SAT predominated over SFA, and PUFA particularly predominated over MUFA. M pod was found to be the richest in all amino acid values except isoleucine, lysine, and valine. The essential and nonessential amino acids in okra are comparable to those in soybeans. Therefore, okra could be used as a good source of proteins for human nutrition. Seeds can be subjected to oil extraction for additional benefits. Okra can be used in making soaps, emulsifiers, and quick-drying oils and has become increasingly popular in the cosmetics industry.


Chorea and convulsive seizures are rare consequences of hyperthyroidism , 2. We d4 concentrations were 0.01 μIU/ml and 43 pmol/l, respectively . The patient was treated with thiamazole in tapering doses (from 60 to 10 mg per day). After 2 months, hypothyreosis was observed and thiamazole was withdrawn. 2 weeks later, the signs of thyrotoxicosis and involuntary movements of the left extremities appeared. Hyperthyreosis was verified (fT4 87.3 pmol/l) and treatment with propylothiouracil (600 mg per day) was initiated. Neurological examination confirmed left-sided hemichorea. Elevated levels of TSH receptor antibodies (TRAb) and thyroperoxidase autoantibodies (TPO Ab) were documented: 16.7 and 257.5 IU/ml, respectively . Biopsy specimen of the thyroid showed lymphocytic infiltration, typical of lymphocytic thyroiditis. The patient had no family history of neurological disorders or dementia syndromes. Ceruloplasmin and copper concentrations were normal, similar to anticardiolipin, antistreptolysin O, and anti-neutrophil cytoplasm antibodies. Antinuclear and anti-double-stranded DNA antibodies were negative. Cerebrospinal fluid examination and MRI of the brain were normal, and borreliosis and toxoplasmosis were ruled out based on immunological examination of serum and cerebrospinal fluid. EEG revealed slight changes in the posterior temporal and occipital leads: theta waves and isolated sharp waves, predominantly in the right hemisphere, along with slight changes of paroxysmal character in the frontal and anterior temporal leads. Haloperidol (6 mg per day) and thioridazin (50 mg per day) were administered. fT4 concentrations diminished (28.7 pmol/l) and choreic movements considerably subsided.In 2003, a 23-year-old woman presented with the first symptoms of hyperthyroidism—palpitations, weight loss and exercise intolerance. TSH and fTTreatment with a maintenance dose of thiamazole was continued. Prednisone  was administered. The dosages of neuroleptics were reduced and finally withdrawn. Choreic movements diminished gradually to disappear completely within 6 weeks. The return of chorea has not been observed so far.4 concentrations were 0.02 μIU/ml and 51.73 pmol/l, respectively. Thiamazole was withdrawn and propylothiouracil  was administered. It was withdrawn in the third trimester, when serum fT4 decreased to 14.7 pmol/l. 3 months after the delivery, signs of hyperthyreosis were observed (fT4 = 41.7 pmol/l), and treatment with propylothiouracil (150 mg/day) was reinstated. In October 2006, the patient presented with generalized tonic–clonic convulsive seizures, loss of consciousness and retrograde amnesia. fT3 and fT4 concentrations were elevated . Laboratory tests and MRI of the brain were normal. Antithyroid treatment was intensified and the patient became euthyroid in December 2006. TPO Ab and anti-thyroglobulin antibodies (Tg Ab) were elevated . In June 2008, the patient underwent treatment with radioactive iodine 131I (150 MBq).For over a year, during treatment with thiamazole (2.5 mg per day), the patient was euthyroid. In October 2005, in the first trimester of pregnancy, symptoms of thyreotoxicosis were once again reported. TSH and fTBetween January 2009 and August 2010, the patient was euthyroid and did not receive any pharmacotherapy. During this period of time, she started to suffer from depression, difficulties with her memory, and sleep disturbances. Psychological examination confirmed mild cognitive impairment and mood disorder. Single-photon emission computed tomography of the brain showed slight global hypoperfusion in both cerebral hemispheres, and an unilateral reduction in the regional blood flow of the left parietal lobe, left temporal lobe and left insula. MRI of the brain was normal. EEG revealed slight changes in frontal and temporal leads in the form of theta waves and isolated sharp waves, accompanied by minor paroxysmal features observed in the posterior temporal and occipital leads.4 1.36 ng/dl) and treatment with l-thyroxin (25 μg/day) was initiated. Cognitive function deterioration has not been observed thus far.In January 2010, levels of TPO Ab were still elevated (324 IU/ml), while TG Ab and TRAb were normal. In August 2010, subclinical hypothyreosis was diagnosed (TSH 6.13 μIU/ml, fTThe pathophysiological mechanism underlying hyperthyroidism-induced chorea is unclear. In most reported cases, choreoathetosis was observed during thyreotoxic states, and one cannot exclude autoimmune mechanisms underlying the chorea , 3. MorpHereby, presented case illustrates that cognitive impairment may be an unusual consequence of autoimmune thyroid disease, and thyroid function should be examined in subjects with encephalopathy, chorea and seizures.


A model reaction, the reduction of styrene to ethylbenzene through a 10% Pd/C catalyst bed, was used to examine performance at various pressure settings, over sequential runs, and with commercial catalyst cartridges. In addition, the consistency of the hydrogen flow was indirectly measured by in-line UV spectroscopy. Finally, system contamination due to catalyst leaching, and the resolution of this issue, is described. The impact of these factors on the run-to-run reproducibility of the H-Cube® reactor for screening and reaction optimization is discussed.The performance of the ThalesNano H-Cube The potential advantages of heterogeneous catalytic flow hydrogenation over traditional batch reactor processes are many and significant . In part®, designed for smaller-scale use in academic and drug discovery labs . An Agilent J&W DB-XLB 30 m × 0.25 mm i.d. capillary column, with a 0.5 micron film, was used in combination with the following oven temperature program: An initial temperature of 70 °C held for 5.0 min, then a 50 °C/min ramp to the final temperature of 200 °C, held for 1.0 min. The injector temperature was set to 250 °C and the ion source temperature was set to 230 °C. Helium  was used as the carrier gas with a gas flow of 37 cm/s average linear velocity and at a pressure of 8.90 psi. Either a split method  or a splitless method was used, depending upon sample concentration. Samples were diluted to 0.04 M when the split method was used, or to 0.004 M when the splitless method was used. Mass data was acquired after a 3.00 min solvent delay, and scan parameters covered the range of 15.0–550.0 amu. Data analysis was performed using Agilent GC/MSD Chemstation software.Supporting information features experimental queues for which an “instability detected” status was recorded by the auto sampler software during the run, and a full data set for the alternating Pd/quartz experiment.File 1Experimental queues and alternating Pd/quartz results.