Effects of replacing fish meal with cottonseed protein concentrate on the growth, immune responses, digestive ability and intestinal microbial flora in Litopenaeus vannamei.

The effects of cottonseed protein concentrate (CPC) in place of fishmeal on the growth performance, immune response, digestive ability and intestinal microbiota of Litopenaeus vannamei were investigated in this study. L. vannamei (initial body weight: 0.42 ± 0.01g) was fed for 8 weeks by four isonitrogenous and isolipid feeds with CPC replacing fishmeal (FM) at 0% (control), 15% (CPC15), 30% (CPC30) and 45% (CPC45), respectively. At the end of the study, the final body weight (FBW), weight gain rate (WGR), specific growth rate (SGR) and protein efficiency ratio (PER) of L. vannamei in CPC15 and CPC30 groups were significantly increased, while the feed conversion ratio (FCR) of L. vannamei in the CPC30 group was significantly reduced when compared with the FM group (P < 0.05). After Vibrio parahaemolyticus infection, the cumulative mortality of L. vannamei in CPC15 within 24 hpi was significantly lower than that of the control group (P < 0.05). When compared with the control group, the activities and expression of the immunity-related enzymes in the hepatopancreas had almost the same obvious change trend in the CPC-containing groups, which indicated that the replacement for fishmeal by CPC led to significant immune response in L. vannamei. Besides, significant up-regulation of the digestive enzyme activities were observed in the CPC-containing groups. Analysis of intestinal microbiota showed that significant difference in alpha diversity existed between the CPC-containing groups and the control group. The relative abundances of several top 10 dominated species at the phylum and genus levels were significantly changed in the CPC-containing groups compared with the control group (P < 0.05). Functional prediction of the microbiota indicated that the pathway of protein digestion and absorption was significantly more abundant while the pathways of nitrotoluene degradation, aminobenzoate degradation, atrazine degradation, dioxin degradation and xylene degradation were significantly less abundant in the CPC-containing groups than the FM group (P < 0.05). In summary, optimal dietary CPC replacement of FM could improve the growth, immunity, digestive capacity and the diversities of the intestinal microbial flora of L. vannamei. However, parts of the functions of the intestinal microbial flora were decline.

EFFECTS OF TRICAINE METHANESULFONATE IN A MANAGED COLLECTION OF MOON JELLYFISH (AURELIA AURITA).

The moon jellyfish (Aurelia aurita) is a scyphozoan frequently maintained in public and private aquaria. Little research has been conducted to investigate the effects of various drugs, such as anesthetics, in this species. Tricaine methanesulfonate (MS-222), a common immersion anesthetic for fish and amphibians, was evaluated in a managed population of moon jellyfish. Twenty-four clinically healthy jellyfish were assigned into three groups of eight for trials of 0.3 g/L MS-222 (low concentration [LC]), 0.6 g/L MS-222 (high concentration [HC]), and a saltwater control. The goal was to evaluate the effects of MS-222 administration on moon jellyfish movement and response to stimuli. Movement and response to stimuli were measured via rocking and probe stimulus tests and observations of bell contraction quality and body tone. These tests were performed at baseline and throughout both drug exposure and recovery periods. A threshold drug effect was defined based on systematic scoring criteria. Additionally, elastomer tags were administered to four of eight animals in each MS-222 group to evaluate response to tag placement after drug exposure. Threshold drug effect was achieved in six of eight individuals in the LC group and eight of eight individuals in the HC group. The LC group had median threshold and recovery times of 12.2 and 10.1 min, respectively, while the HC group had median threshold and recovery times of 4.0 and 19.9 min, respectively. The HC group had significantly faster time to threshold drug effect (P < 0.001) and longer recovery times (P= 0.005) than the LC group. In both the LC and HC tagged group, three of four jellyfish had no reaction to tag placement. All animals recovered uneventfully, and there were no mortalities. MS-222 at 0.3 and 0.6 g/L decreased movement and response to stimuli in moon jellyfish.

USE OF TRICAINE METHANESULFONATE, CLOVE OIL AND 2-PHENOXYETHANOL FOR IMMERSION EUTHANASIA OF UNGA CICHLIDS (SAROTHERODON LINELLII).

Fish may warrant euthanasia for a variety of reasons, but euthanasia may be difficult to accomplish or confirm because fish can recover from deep anesthesia even after cardiac and respiratory arrest. The efficacies of three types of anesthetics were evaluated to compare their suitability for euthanasia of Unga cichlids (Sarotherodon linellii). Thirty cichlids were randomly assigned to be immersed in one of the three anesthetic solutions: tricaine methanesulfonate (MS-222), 2-phenoxyethanol (2-PE), and clove oil (CO) at doses of 1,000 mg/L, 2 mL/L, and 500 mg/L respectively. The opercular rates and caudal fin stroke rates were quantified, and the time to cessation of physiological measures (CPM) including caudal fin strokes, the reaction to external stimuli, the righting reflex, swimming, and operculation were recorded. Varying anesthetic induction times were observed with all three euthanasia solutions; the time to CPM in the 2-PE group occurred at a significantly slower rate than in the MS-222 group (P < 0.01). No significant differences were identified for the time to CPM when comparing the standard length or weight of the cichlids in all euthanasia solutions (P > 0.05). The cost of euthanasia per cichlid was calculated, with the most economically viable option being 2-PE; at more than seven times the price of 2-PE, MS-222 was the most expensive. After a 60-min immersion in the euthanasia solution, the presence of an audible heartbeat was identified in 100% of the cichlids immersed in 2-PE, 100% immersed in CO, and 90% in MS-222, indicating that they were not reliably euthanized. Therefore, a two-step protocol is recommended in cichlids for euthanasia: heavy anesthesia via immersion followed by an intravenous or intracardiac injection of euthanasia solution, or other secondary method of euthanasia.

The effects of commonly used anaesthetics on colour measurements across body regions in the poeciliid fish, Girardinus metallicus.

The effects of common anaesthetics on the hue, saturation and brightness measurements of the poeciliid fish Girardinus metallicus were investigated in two experiments. For both experiments the coloration of four body regions was measured from digital images of the same males obtained under three conditions: (1) control (in a water-filled chamber); (2) anaesthetised with MS-222; and (3) anaesthetised with eugenol (clove oil). In experiment 1 anaesthetised fish were photographed out of water. In experiment 2 all photographs were taken in a water-filled chamber. Anaesthetics altered coloration in both experiments. In the more methodologically consistent experiment 2 we found significantly different hue, increased saturation and decreased brightness in anaesthetic v. control conditions, consistent with darkening caused by the anaesthetics. The body regions differed in coloration consistent with countershading but did not differentially change in response to anaesthesia. These findings suggest that photographing fish in a water-filled chamber without anaesthetic is preferable for obtaining digital images for colour analysis and that multiple body regions of fish should be measured when assessing coloration patterns meaningful in behavioural contexts, to account for the gradients caused by countershading. We are encouraged that some researchers employ such methods already and caution against using anaesthetics except when absolutely necessary for immobilisation.

Modulation of rainbow trout (Oncorhynchus mykiss) cutaneous mucosal immune responses following anesthesia: A comparative study on different anesthetic agents.

The present study investigated the possible effects of different anesthetic agents including MS222 (50 ppm), 2-Phenoxyethanol (2-PE) (0.2 mL L-1) and clove oil (25 ppm), on cutaneous mucosal immune parameters in rainbow trout (Oncorhynchus mykiss). The induction and recovery times for each anesthetic agent were assessed. Also, the immune parameters were measured in skin mucus, 1 and 24 h post anesthesia. No significant difference was observed among treatments at 1 h post-anesthesia except for bactericidal and alkaline phosphatase (ALP) activities which was significantly enhanced in fish exposed to 2-PE compared to other anesthetics. At 24 h post-anesthesia, most of the skin mucosal immune parameters were increased upon exposure to clove oil but decreased following exposure to 2-PE. However, no significant change was noticed after MS222 exposure. These results demonstrated that the anesthetics type should be considered to avoid possible immunosuppression in farmed fish. Furthermore, the present results could be useful for better understanding of alterations in cutaneous mucosal immunity in response to chemical stressors such as anesthetic agents.

Anesthetic protocol for microinjection-related handling of Siberian sturgeon (Acipenser baerii; Acipenseriformes) prolarvae.

An anesthetic protocol was optimized for microinjection-related handling of Siberian sturgeon (Acipenser baerii; Acipenseriformes) prolarvae, an extant primitive fish species commonly grown in aquaculture. Comparative examinations of three selected anesthetics (clove oil, lidocaine, and MS-222) with a dosage regime of 50, 100, 200, and 400 mg/L indicated that MS-222 was the most efficient agent for Siberian sturgeon prolarvae, as evidenced by the fast induction of anesthesia with quick and uniform recovery. Meanwhile, clove oil should be avoided, due to prolonged recovery times varying widely between individuals. None of the tested anesthetics significantly affected prolarval viability at any of the dosage regimes tested in this study. Based on an analysis of the duration of an unconscious state in air, we recommend a dose of 200 mg/L MS-222 for microinjection. Recovery time after use of this dose was influenced by the prolarval age and the development of gills, in which prolarvae older than 3 days after hatching required longer recovery times than did younger prolarvae. Post-recovery behavioral assessment showed no apparent difference between MS-222-anesthetized and non-anesthetized prolarvae in their swimming behavior and phototactic responses. Applicability of currently developed anesthetic protocol using MS-222 in larval microinjection was demonstrated with the injection of a visible dye to the anesthetized prolarvae, followed by the analysis of post-recovery viability. Taken together, the present anesthetic protocol based on 200 mg/L of MS-222 could provide researchers with practical usefulness with good safety margins for the micromanipulation and other related handlings of Siberian sturgeon prolarvae.

Effect of the Level of Anesthesia on the Auditory Brainstem Response in the Emei Music Frog (Babina daunchina).

Anesthesia is known to affect the auditory brainstem response (ABR) in mice, rats, birds and lizards. The present study investigated how the level of anesthesia affects ABR recordings in an amphibian species, Babina daunchina. To do this, we compared ABRs evoked by tone pip stimuli recorded from 35 frogs when Tricaine methane sulphonate (MS-222) anesthetic immersion times varied from 0, 5 and 10 minutes after anesthesia induction at sound frequencies between 0.5 and 6 kHz. ABR thresholds increased significantly with immersion time across the 0.5 kHz to 2.5 kHz frequency range, which is the most sensitive frequency range for hearing and the main frequency range of male calls. There were no significant differences for anesthetic levels across the 3 kHz to 6 kHz range. ABR latency was significantly longer in the 10 min group than in the 0 and 5 min groups at frequencies of 0.5, 1.0, 1.5, 2.5 kHz, while ABR latency did not differ across the 3 kHz to 4 kHz range and at 2.0 kHz. Taken together, these results show that the level of anesthesia affects the amplitude, threshold and latency of ABRs in frogs.

Sex differences and endocrine regulation of auditory-evoked, neural responses in African clawed frogs (Xenopus).

Mating depends on the accurate detection of signals that convey species identity and reproductive state. In African clawed frogs, Xenopus, this information is conveyed by vocal signals that differ in temporal patterns and spectral features between sexes and across species. We characterized spectral sensitivity using auditory-evoked potentials (AEPs), commonly known as the auditory brainstem response, in males and females of four Xenopus species. In female X. amieti, X. petersii, and X. laevis, peripheral auditory sensitivity to their species own dyad-two, species-specific dominant frequencies in the male advertisement call-is enhanced relative to males. Males were most sensitive to lower frequencies including those in the male-directed release calls. Frequency sensitivity was influenced by endocrine state; ovariectomized females had male-like auditory tuning while dihydrotestosterone-treated, ovariectomized females maintained female-like tuning. Thus, adult, female Xenopus demonstrate an endocrine-dependent sensitivity to the spectral features of conspecific male advertisement calls that could facilitate mating. Xenopus AEPs resemble those of other species in stimulus and level dependence, and in sensitivity to anesthetic (MS222). AEPs were correlated with body size and sex within some species. A frequency following response, probably encoded by the amphibian papilla, might facilitate dyad source localization via interaural time differences.

Crystal structures of the pyrazinamide-p-aminobenzoic acid (1/1) cocrystal and the transamidation reaction product 4-(pyrazine-2-carboxamido)benzoic acid in the molten state.

The synthesis of pharmaceutical cocrystals is a strategy to enhance the performance of active pharmaceutical ingredients (APIs) without affecting their therapeutic efficiency. The 1:1 pharmaceutical cocrystal of the antituberculosis drug pyrazinamide (PZA) and the cocrystal former p-aminobenzoic acid (p-ABA), C7H7NO2·C5H5N3O, (1), was synthesized successfully and characterized by relevant solid-state characterization methods. The cocrystal crystallizes in the monoclinic space group P21/n containing one molecule of each component. Both molecules associate via intermolecular O-H···O and N-H···O hydrogen bonds [O···O = 2.6102 (15) Šand O-H···O = 168.3 (19)°; N···O = 2.9259 (18) Šand N-H···O = 167.7 (16)°] to generate a dimeric acid-amide synthon. Neighbouring dimers are linked centrosymmetrically through N-H···O interactions [N···O = 3.1201 (18) Šand N-H···O = 136.9 (14)°] to form a tetrameric assembly supplemented by C-H···N interactions [C···N = 3.5277 (19) Šand C-H···N = 147°]. Linking of these tetrameric assemblies through N-H···O [N···O = 3.3026 (19) Šand N-H···O = 143.1 (17)°], N-H···N [N···N = 3.221 (2) Šand N-H···N = 177.9 (17)°] and C-H···O [C···O = 3.5354 (18) Šand C-H···O = 152°] interactions creates the two-dimensional packing. Recrystallization of the cocrystals from the molten state revealed the formation of 4-(pyrazine-2-carboxamido)benzoic acid, C12H9N3O3, (2), through a transamidation reaction between PZA and p-ABA. Carboxamide (2) crystallizes in the triclinic space group P1̅ with one molecule in the asymmetric unit. Molecules of (2) form a centrosymmetric dimeric homosynthon through an acid-acid O-H···O hydrogen bond [O···O = 2.666 (3) Šand O-H···O = 178 (4)°]. Neighbouring assemblies are connected centrosymmetrically via a C-H···N interaction [C···N = 3.365 (3) Šand C-H···N = 142°] engaging the pyrazine groups to generate a linear chain. Adjacent chains are connected loosely via C-H···O interactions [C···O = 3.212 (3) Šand C-H···O = 149°] to generate a two-dimensional sheet structure. Closely associated two-dimensional sheets in both compounds are stacked via aromatic π-stacking interactions engaging the pyrazine and benzene rings to create a three-dimensional multi-stack structure.

Comparison of the effect of four anaesthetics on haematological profiles, oxidative stress and antioxidant enzymes in barbel (Barbus barbus).

OBJECTIVES: Currently, many questions regarding the effect of anaesthetics to fish remain unresolved. Fish species may differ widely in their response to an anaesthetic, the screening of dosages is often necessary. The aim of this study was to compare the effect of tricaine methane sulphonate (MS 222), clove oil, 2-phenoxyethanol and Propiscin on haematological profiles, oxidative stress biomarkers and antioxidant enzymes in barbel (Barbus barbus). DESIGN: The haematological profiles, oxidative stress biomarkers and antioxidant enzymes of barbel were evaluated immediately after a 10 min anaesthesia (MS 222--100 mg.L(-1), clove oil--33 mg.L(-1), 2-phenoxyethanol--0.4 mg.L(-1), Propiscin--1.0 mg.L(-1)), and 24 h after anaesthesia. RESULTS: The 10 min exposure in the recommended concentrations of tested anaesthetics have no significant effect on haematological profiles, levels of thiobarbituric acid reactive substances, and activity of glutathione reductase of barbel. The activity of superoxide dismutase (SOD) was significantly decreased (p<0.01) in the muscle in all experimental groups. The activity of SOD showed a significant decrease (p<0.01) in the liver 24 h after all anaesthetics; however in the gill the activity of SOD was significantly increased (p<0.01) in Propiscin (10 min). The activity of catalase (CAT) was significantly increased (p<0.05) in the muscle 24 h after all anaesthetics. CONCLUSIONS: The observed effects on barbel antioxidant systems may be a defence against oxidative damage. The results of this study suggest that the antioxidant systems of barbel are altered by Propiscin anaesthesia, but are slightly affected by MS 222, clove oil, and 2-phenoxyethanol anaesthesia.

Conditioned place avoidance of zebrafish (Danio rerio) to three chemicals used for euthanasia and anaesthesia.

Zebrafish are becoming one of the most used vertebrates in developmental and biomedical research. Fish are commonly killed at the end of an experiment with an overdose of tricaine methanesulfonate (TMS, also known as MS-222), but to date little research has assessed if exposure to this or other agents qualifies as euthanasia (i.e. a "good death"). Alternative agents include metomidate hydrochloride and clove oil. We use a conditioned place avoidance paradigm to compare aversion to TMS, clove oil, and metomidate hydrochloride. Zebrafish (n = 51) were exposed to the different anaesthetics in the initially preferred side of a light/dark box. After exposure to TMS zebrafish spent less time in their previously preferred side; aversion was less pronounced following exposure to metomidate hydrochloride and clove oil. Nine of 17 fish exposed to TMS chose not to re-enter the previously preferred side, versus 2 of 18 and 3 of 16 refusals for metomidate hydrochloride and clove oil, respectively. We conclude that metomidate hydrochloride and clove oil are less aversive than TMS and that these agents be used as humane alternatives to TMS for killing zebrafish.

Effect of vanillic acid on COQ6 mutants identified in patients with coenzyme Q10 deficiency.

Human COQ6 encodes a monooxygenase which is responsible for the C5-hydroxylation of the quinone ring of coenzyme Q (CoQ). Mutations in COQ6 cause primary CoQ deficiency, a condition responsive to oral CoQ10 supplementation. Treatment is however still problematic given the poor bioavailability of CoQ10. We employed S. cerevisiae lacking the orthologous gene to characterize the two different human COQ6 isoforms and the mutations found in patients. COQ6 isoform a can partially complement the defective yeast, while isoform b, which lacks part of the FAD-binding domain, is inactive but partially stable, and could have a regulatory/inhibitory function in CoQ10 biosynthesis. Most mutations identified in patients, including the frameshift Q461fs478X mutation, retain residual enzymatic activity, and all patients carry at least one hypomorphic allele, confirming that the complete block of CoQ biosynthesis is lethal. These mutants are also partially stable and allow the assembly of the CoQ biosynthetic complex. In fact treatment with two hydroxylated analogues of 4-hydroxybenzoic acid, namely, vanillic acid or 3-4-hydroxybenzoic acid, restored the respiratory growth of yeast Δcoq6 cells expressing the mutant huCOQ6-isoa proteins. These compounds, and particularly vanillic acid, could therefore represent an interesting therapeutic option for COQ6 patients.

Efficacy of different anaesthetics for pikeperch (Sander lucioperca L.) in relation to water temperature.

OBJECTIVES: The objectives of the study were to compare the different doses of clove oil, Propiscin, and tricaine methane sulphonate (MS 222) in relation to water temperature in pikeperch aquaculture. DESIGN: For assessment of this experiment 168 fish (10.77 ± 0.59 cm total body length and 7.88 ± 1.74 g body weight) were used. Three different anaesthetic treatments (Propiscin, clove oil and MS 222) were used. Three doses of each anaesthetic treatment (Propiscin: 0.5; 1; 1.5 ml x L(-1), clove oil: 15; 30; 60 mg x L(-1), MS 222: 50; 100; 150 mg x L(-1)) were compared at three different temperatures 9.5; 15.5 and 23 degrees C. RESULTS: In comparison of these doses of anaesthetic in different temperature, the significantly shortest time to attain phase A7 (total complete anaesthesia) was observed for Propiscin (1.5 ml L(-1)) 0:31 ± 0:04 min (23 degrees C) to 0:33 ± 0:25 min (9.5 degrees C) compared to MS 222 (150 mg x L(-1)) 1:04 ± 0:21 min (23 degrees C) to 1:54 ± 0:32 min (9.5 degrees C) and clove oil (60 mg x L(-1)) 1:05 ± 0:17 min (23 degrees C) to 3:05 ± 0:31 min (9.5 degrees C). On the other hand, the longest time of anaesthesia recovery was attained using Propiscin (1.5 ml x L(-1)) 10:35 ± 1:40 min (23 degrees C) to 32:30 ± 1:10 min (9.5 degrees C) compared to clove oil (60 mg x L(-1)) 2:39 ± 0:50 min (23 degrees C) to 9:36 ± 2:34 min (60 mg x L(-1), 9.5 degrees C) and MS 222 (150 mg x L(-1)) 2:26 ± 1:27 min (23 degrees C) to 4:59 ± 0:39 min (9.5 degrees C). CONCLUSIONS: The results from this study showed that the optimal and sufficient doses in all tested temperatures for pikeperch are 30 mg x L(-1) of clove oil, 100 mg x L(-1) of MS 222 and 0.5 ml x L(-1) of Propiscin.

Effect of tricainemethanesulfonate (MS222), clove oil and electro-anaesthesia on respiratory burst activity in whole blood and serum alternative complement response in rainbow trout (Oncorhynchus mykiss), during the narcosis stage.

There is a little available information on the suppressive effect of anaesthesia on immune response in fish, especially electro-anaesthesia. In the present study, two anaesthetics, MS222 (50 ppm), clove oil (25 ppm), and electro-anaesthesia were tested in rainbow trout (Oncorhynchus mykiss) during the narcosis stage in order to observe their effects on the innate immune system. The results showed that electro-anaesthesia reduces light emission in chemiluminescence assay both 1 and 24 h post anaesthesia. Clove oil and MS222 decreased light emission 24 h post anaesthesia. In addition, clove oil, MS222 and electro-anaesthesia had no effect on alternative complement (ACH50) response. From the perspective of aquaculture practice, these data show that the type of anaesthesia should be taken into account to avoid possible immunosuppression in rainbow trout.

The effect of three anaesthetic protocols on the stress response in cane toads (Rhinella marina).

OBJECTIVE: Three anaesthetics (MS222, clove oil and a mixture of ketamine/diazepam) were administered to cane toads to determine their effect on the hypothalamic-pituitary-adrenal (HPA) axis. Time to induction and recovery and any adverse events were also evaluated. STUDY DESIGN: Prospective randomized experimental trial. ANIMALS: Thirty adult male cane toads (Rhinella marina) with body mass ranging between 130 and 250 g were captured from the field. METHODS: Three groups of 10 toads were anaesthetized with ketamine (200 mg kg(-1) ) and diazepam (0.2 mg kg(-1) ) by intramuscular injection, MS222 (3 g L(-1) ) or clove-oil (0.3 mL L(-1) ) both by immersion. Blood samples were collected to determine plasma corticosterone concentrations. Induction and recovery time were recorded in each treatment. After full recovery animals were euthanized and a complete post-mortem examination was performed. RESULTS: Significant differences were found in the activation of the HPA axis and in the times of induction and recovery between treatments (p < 0.001). Animals anaesthetized with clove-oil had the highest levels of corticosterone in plasma (42.5 ± 21.6 ng mL(-1) ). No differences were found between ketamine/diazepam (15.0 ± 13.3 ng mL(-1) ) and MS222 (22.0 ± 13.6 ng mL(-1) ) groups. The mean ± SD induction (minutes) and recovery (hours) times respectively were; ketamine/diazepam 66.5 ± 11 and 8 ± 3, clove oil 39 ± 12 and 7.6 ± 3, and MS222 42.5 ± 11 and 1.5 ± 0.5. Clove oil exposure had 30% mortality. Death followed a period of respiratory distress with changes consistent with non-cardiogenic oedema observed at post-mortem examination. CONCLUSIONS AND CLINICAL RELEVANCE: Based on shorter induction and recovery times and minimal activation of HPA, MS222 is the anaesthetic of choice in cane toads. If it is not possible to use immersion methods of anaesthesia, ketamine/diazepam can be used but induction and recovery times are prolonged. Clove oil had unacceptable mortality in this study and should be used with extreme caution.

[Synthesis of 2-hydroxyl-5-butyramidobenzoic acid and its effect on acetic acid-induced colitis in rats].

OBJECTIVE: To study the method for synthesis of 2-hydroxyl-5- butyramidobenzoic acid and test its effect on acetic acid-induced colitis in rats. METHODS: 2-hydroxyl-5-butyramidobenzoic acid was synthesized from 5-aminosalicylic acid and butyric acid by amidation, esterification and hydrolization. The effect of 2-hydroxyl-5-butyramidobenzoic acid on acetic acid enema-induced colitis in rats was investigated. RESULTS: The structure of 2-hydroxyl-5-butyramidobenzoic acid was identified by IR and 1H-NMR. After treatment with acetic acid, the colon mucosal damage index (CMDI), fecal occult blood (OB) test, and activity of myelperoxidase (MPO) increased significantly in the rats as compared to the control levels. 2-hydroxyl-5- butyramidobenzoic acid obviously reduced the CMDI and OB, and reduced the level of MPO in the rats with colitis. CONCLUSION: The synthesis of 2-hydroxyl-5-butyramidobenzoic acid requires only mild conditions with simple procedures, and the synthesized 2-hydroxyl-5-butyramidobenzoic acid shows obvious therapeutic effects on mucosal damage of in rats with acetic acid-induced colitis.

Synthesis and anti-platelet evaluation of 2-benzoylaminobenzoate analogs.

Fifty-two 2-benzoylaminobenzoate analogs were synthesized and subjected to anti-platelet aggregation assay using arachidonic acid (AA), collagen (Col), thrombin (Thr), and U46619 as inducers. The results revealed that most of 2-benzoylaminobenzoic acid derivatives showed a selectively inhibitory effect on AA-induced platelet aggregation. As a result of the 2-benzoylaminobenzoic acid derivatives (18, 44, and 46), there were no inhibitory effects on platelet aggregation induced by U46619, but these elicited an inhibitory effect on thromboxane B(2) formation at 1.0microM. These 2-benzoylaminobenzoate analogs were therefore proposed as cyclooxygenase inhibitors.

Screening of telomerase inhibitors.

Shortening of telomeres prevents cells from uncontrolled proliferation. Progressive telomere shortening occurs at each cell division until a critical telomeric length is reached. Telomerase expression is switched off after embryonic differentiation in most normal cells, but it is expressed in a very high percentage of tumors of different origin. Thus, telomerase is regarded as the best tumor marker and a promising novel molecular target for cancer treatment. Therefore, different strategies to inhibit telomerase have been developed. However, systematic screening of telomerase inhibitors has not been performed to compare their therapeutic potential. We propose a suitable strategy for estimation of the therapeutic potential of telomerase inhibitors, which is based on a systematic screening of different inhibitors in the same cell system. From the long list of compounds discussed in the literature, we have selected four telomerase inhibitors of different structure and mode of action: BRACO19 (G-quadruplex-interactive compound), BIBR1532 (non-nucleosidic reverse transcriptase inhibitor), 2'-O-methyl RNA, and peptide nucleic acids (PNAs; hTR antisense oligonucleotides). To determine minimal effective concentrations for telomerase inhibition, telomerase activity was measured using the cell-free telomerase repeat amplification protocol (TRAP) assay. We also tested inhibitors in long-term cell-culture experiments by exposing A-549 cells to non-cytotoxic concentrations of inhibitors for a period of 99 days. Subsequently, telomerase activity of A-549 cells was investigated using the TRAP assay, and telomere length of samples was assessed by telomere restriction fragment (TRF) Southern blot analysis.

Differential effects of anesthetics on electrical properties of the rainbow trout (Oncorhynchus mykiss) heart.

We compared electrocardiographic signals in hatchery-reared, non-spinally-transected, immature rainbow trout (Oncorhynchus mykiss Walbaum) under clove oil (25 ppm), tricaine methanesulfonate (tricaine, 60 ppm), and benzocaine (108 ppm) general anesthesia (35 min, 14 degrees C). For all 3 anesthetics, the mean heart rate (HR) and QRS amplitude did not differ, and QRS duration and QT interval were independent of HR. Heart rate variability (HRV) was significantly (4-fold, P=0.032) higher under benzocaine than under clove oil and tricaine, but did not differ between clove oil and tricaine. QRS duration differed between groups (P<0.001, F=121); benzocaine anesthesia resulted in longer QRS complexes compared to clove oil (P<0.001) and tricaine (P<0.001) anesthesia, and QRS complexes under clove oil were longer than those under tricaine (P<0.001). High HRV and QRS amplitude variation with benzocaine were associated with HR oscillations as anesthetic exposure time increased, and suggest benzocaine toxicity which may influence cardiac function studies. Similar clove oil and tricaine ECG patterns suggest comparable autonomic effects, and maintenance of myocardial excitability. Given its low cost, ease of use, and similar ECG profiles to tricaine, clove oil is a viable alternative for studies of cardiac function in anesthetized rainbow trout.

Regulation and expression of heme oxygenase enzymes in aged-rat brain: age related depression in HO-1 and HO-2 expression and altered stress-response.

The heme oxygenase isozymes, HO-1 and HO-2, oxidatively cleave the heme molecule to produce biliverdin and the gaseous messenger, CO. The cleavage results in the release of iron, a regulator of transferrin, ferritin, and nitric oxide (NO) synthase gene expression. Biliverdin reductase (BVR) then catalyzes the reduction of biliverdin, generating the potent intracellular antioxidant, bilirubin. We report an age-related decrease in HO-1 and HO-2 expression present in select brain regions including the hippocampus and the substantia nigra, that are involved in the high order cognitive processes of learning and memory. The age-related loss of monoxide-producing potential in select regions of the brain was not specific to the HO system but was also observed in neuronal NO-generating system. Furthermore, compared to 2-month old rats, the ability of aged brain tissue to respond to hypoxic/hyperthermia was compromised at both the protein and the transcription levels as judged by attenuated induction of HO-1 immunoreactive protein and its 1.8 Kb transcript. Neotrofin (AIT), a cognitive-enhancing and neuroprotective drug, caused a robust increase in HO-1 immunoreactive protein in select neuronal regions and increased the expression of HO-2 transcripts. The potential interplay between regulation of HO-2 gene expression and the serum levels of the adrenal steroids is discussed. We suggest the search for therapeutic agents that reverse the decline and aberrant stress response of HO enzymes may lead to effective treatment regimens for age-associated neuronal deficits.