THE USE OF BENZOCAINE HYDROCHLORIDE AS FISH TRANQUILLIZER AND ANAESTHETIC IN SALINE WATERS

SUMMARY

The anaesthetic effects of various concentrations of benzocaine hydrochloride were tested on Liza macrolepis and Sarotherodor mossambicus in sea water and diluted sea water, respectively. Induction time for anaesthesia was negatively correlated with increasing anaesthetic concentrations in L. macrolepis.

In S. mossambicus, however, operculum clamping appeared to be responsible-f or induction times increasing with increased anaesthetic concentration.

The tranquillizing effects of low concentrations of benzocaine hydrochloride on L. macrolepis was also studied.     

The uptake of the anaesthetic benzocaine hydrochloride by the gills and the skin of three freshwater fish species

The uptake of benzocaine hydrochloride and neutralized benzocaine hydrochloride by the skin and the gills of Cyprinus carpio, Oreochromis mossambicus and Salmo gairdneri were studied. The differences observed can mainly be ascribed to degree of ionization and the lipid solubility of the anaesthetic.     

Breath holding during intense exercise: arterial blood gases, pH, and lactate

Seven healthy endurance-trained [maximal O2 uptake (VO2max) = 57.1 +/- 4.1 ml.kg-1.min-1)] female volunteers (mean age 24.4 +/- 3.6 yr) served as subjects in an experiment measuring arterial blood gases, acid-base status, and lactate changes while breath holding (BH) during intense intermittent exercise. By the use of a counterbalance design, each subject repeated five intervals of a 15-s on:30-s off treadmill run at 125% VO2max while BH and while breathing freely (NBH). Arterial blood for pH, PO2, PCO2, O2 saturation (SO2) HCO3, and lactate was sampled from a radial arterial catheter at the end of each work and rest interval and throughout recovery, and the results were analyzed using repeated-measures analysis of variance. Significant reductions in pHa (delta mean = 0.07, P less than 0.01), arterial PO2 (delta mean = 24.2 Torr, P less than 0.01), and O2 saturation (delta mean = 4.6%, P less than 0.01) and elevations in arterial PCO2 (delta mean = 8.2 Torr, P less than 0.01) and arterial HCO3 (delta mean = 1.3 meq/l, P = 0.05) were found at the end of each exercise interval in the BH condition. All of the observed changes in arterial blood gases and acid-base status induced by BH were reversed during the rest intervals. During recovery, significantly (P less than 0.025) greater levels of arterial lactate were found in the BH condition.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of benzocaine on the kinetics of normal and batrachotoxin-modified Na channels in frog node of Ranvier.

The effects of benzocaine (0.5-1 mM) on normal Na currents, and on Na current and gating charge movement (Q) of batrachotoxin (BTX)-modified Na channels were analyzed in voltage-clamped frog node of Ranvier. Without BTX treatment the decay of Na current during pulses to between -40 and 0 mV could be decomposed into two exponential components both in the absence and in the presence of benzocaine. Benzocaine did not significantly alter the inactivation time constant of either component, but reduced both their amplitudes. The amplitude of the slow inactivating component was more decreased by benzocaine than the amplitude of the fast one, leading to an apparently faster decline of the overall Na current. After removal of Na inactivation and charge movement immobilization by BTX, benzocaine decreased the amplitude of INa with no change in time course. INa, QON, and QOFF were all reduced by the same factor. The results suggest that the rate of reaction of benzocaine with its receptor is slow compared to the rates of channel activation and inactivation. The differential effects of benzocaine on the two components of Na current inactivation in normal channels can be explained assuming two types of channel with different rates of inactivation and different affinities for the drug.     

A multiparametric analysis of the synergistic impact of anti-Parkinson's drugs on the fibrillation of human serum albumin

Protein aggregation have been associated with several human neurodegenerative diseases, such as Parkinson's and Alzheimer's diseases. There are several small molecules that can reduce aggregation of proteins. The present study aimed to test the hypothesis that the application of more than one inhibitor either simultaneously or consecutively may result in more efficient inhibition of protein aggregation. To this end, the anti-amyloidogenic behaviour of benserazide hydrochloride (BH) and levodopa (LD) individually and in combination (BH + LD) was investigated using various biophysical, microscopic, and computational techniques. BH, LD, and BH + LD treatments showed inhibitory effects on protein aggregation and had the ability to minimise the amyloid-induced cytotoxicity in human neuroblastoma cell line (SH-SY5Y). The two drugs in combination showed synergism (combination index, CI < 1) between them. These drugs also destabilised the preformed fibrils of human serum albumin (HSA). Our studies consistently showed that the BH + LD treatment showed highest efficacy towards inhibition and disaggregation of amyloid fibrils in comparison to treatment with BH and LD individually. Therefore, application of drugs in combination against fibrillogenesis may represent a new route for development of means for prevention or delaying of the aggregation-related diseases.     

Metamorphosis and the steady state anesthetic concentrations of tricaine,benzocaine and ethanol

In the classical tadpole assay employed for Meyer-Overton type correlations, tricaine and benzocaine are 7–8 times more potent than n-alkanols of equivalent lipid solubility. Median anesthetic concentrations for loss of the righting reflex, AC₅₀(RR)s, were 0.165 and 0.103 mM for tricaine and benzocaine, and log Ps (octanol: water) were 1.81 and 1.98, respectively. Tadpoles receiving a half AC₅₀(RR) each of tricaine and ethanol showed less than additive effects, suggesting a substantial difference in their mechanism(s) of action. AC₅₀(RR)s for both tricaine and benzocaine increased two fold coincident with metamorphosis, reflecting a change from an equilibrium to a nonequilibrium steady state.     

Effects of the local anesthetic benzocaine on the human erythrocyte membrane and molecular models

The interaction of the local anesthetic benzocaine with the human erythrocyte membrane and molecular models is described. The latter consisted of isolated unsealed human erythrocyte membranes (IUM), large unilamellar vesicles (LUV) of dimyristoylphospatidylcholine (DMPC), and phospholipid multilayers of DMPC and dimyristoylphospatidyletanolamine (DMPE), representatives of phospholipid classes located in the outer and inner monolayers of the human erythrocyte membrane, respectively. Optical and scanning electron microscopy of human erythrocytes revealed that benzocaine induced the formation of echinocytes. Experiments performed on IUM and DMPC LUV by fluorescence spectroscopy showed that benzocaine interacted with the phospholipid bilayer polar groups and hydrophobic acyl chains. X-ray diffraction analysis of DMPC confirmed these results and showed that benzocaine had no effects on DMPE. The effect on sodium transport was also studied using the isolated toad skin. Electrophysiological measurements indicated a significant decrease in the potential difference (PD) and in the short-circuit current (Isc) after the application of benzocaine, reflecting inhibition of active ion transport.     

Enhancing the Antitumor Activity of Berberine Hydrochloride by Solid Lipid Nanoparticle Encapsulation

Berberine hydrochloride (BH) is an isoquinolin alkaloid with promising anticancer efficacies. Nevertheless, further development and application of this compound had been hampered by its poor aqueous solubility, low gastrointestinal absorption, and rapid metabolism in the body. In this study, a solid lipid nanoparticle (SLN)-based system was developed for efficient incorporation and persistent release of BH. The drug-loading SLNs (BH-loaded SLNs) were stable, with a mean particle size of 81.42 ± 8.48 nm and zeta potential of −28.67 ± 0.71 mV. BH-loaded SLNs showed desirable drug entrapment efficiency and drug-loaded, and the release of BH from SLNs was significantly slower than free BH. Importantly, our in vitro study indicated that BH-loaded SLNs more significantly inhibited cell proliferation on MCF-7, HepG 2, and A549 cancer cells. Meanwhile, clone formation, cellular uptake, cell cycle arrest, and cell apoptosis studies also demonstrated that BH-loaded SLNs enhanced the antitumor efficacies of BH on MCF-7 cancer cells. Taken together, our results suggest that this SLN formulation may serve as a novel, simple, and efficient system for the delivery of BH.KEY WORDS: antitumor evaluation, apoptosis, berberine hydrochloride, solid lipid nanoparticles     

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.     

Anesthesia of fish with benzocaine does not interfere with comet assay results

Fish blood erythrocytes are frequently used as sentinels in biomonitoring studies. Usually, fish blood is collected by painful cardiac or caudal vein punctures. Previous anesthesia could decrease animal suffering but it is not known at present whether anesthesia can cause confounding effects. Therefore, using the alkaline single cell gel (SCG)/comet assay with blood erythrocytes of the cichlid fish Nile tilapia, we tested for a possible modulation of induced DNA damage (methyl methanesulfonate; MMS) by the anesthetic benzocaine administered by bath exposure (80mg/l for approximately 10min). Furthermore, benzocaine (80-600mg/l) was tested for its genotoxic potential on fish erythrocytes in vitro and for potential interactions with two known genotoxins (MMS and hydrogen peroxide). Our results did neither indicate a significant increase in the amount of DNA damage (even after a 48h follow-up), nor indicated interactions with MMS-induced DNA damage when fish were exposed to benzocaine in vivo. There was also no increase in DNA damage after in vitro exposure of fish erythrocytes to benzocaine. Clear concentration-related effects were observed for the two genotoxins in vitro, which were not significantly altered by the presence of benzocaine. These results suggest that anesthesia of fish does not confound comet assay results and the use of blood samples from anesthetized fish can be recommended with regard to animal welfare.     

The effect of the anaesthetic benzocaine hydrochloride on red cell fragilities in Cyprinus carpio

The effects of benzocaine HCl and neutralized benzocaine HCl anaesthesia were studied on osmotic fragilities in Cyprinus carpio . The differences observed in corpuscular fragility are ascribed to temperatures and pH effects. Further evaluation of such effects are required.     

In vivo effects of the anesthetic,benzocaine, on liver microsomal cytochrome P450 and mixed-function oxidase activities of gilthead seabream (Sparus aurata)

Gilthead seabreams were exposed to benzocaine, 4-aminobenzoic acid ethyl ester, 57 mg/l in sea water for 3 min, daily, for 2 or 3 consecutive days. The fish were killed 20 hr after the last treatment. Benzocaine treatment for 2 or 3 days resulted in 57% and 67% inhibition of liver microsomal aniline 4-hydroxylase and ethylmorphine N-demethylase activities,respectively. The total cytochrome P450 content of fish liver microsomes was unaltered following the 2-day benzocaine treatment. However, additional 3 min benzocaine treatment on day 3 reduced cytochrome P450 level by 50%. Benzocaine produced type II difference spectra with rabbit liver microsomes. Difference spectra of fish liver microsomes elicited by benzocaine were complex. The position of peak and intensity were greatly influenced by the concentration of benzocaine.     

The effects of observer presence on the behavior of Cebus capucinus in Costa Rica

We report on the responses of Cebus capucinus in the Santa Rosa Sector of the Area de Conservación Guanacaste, Costa Rica, to the presence of observers over a 4-week period. Study groups were habituated to different degrees: (1) Cerco de Piedra (CP): continuous observations began in 1984; (2) Exclosure (EX): focus of an 18-month study on males from 1998 to 1999; and (3) NBH: never studied/followed but the group frequently encounters researchers. We collected three types of data: group scans (group state was coded as calm or agitated at observer presence), focal animal data (observer-directed behaviors were recorded), and fecal cortisol levels. The two less-habituated groups (NBH and EX) differed significantly from the habituated group (CP) in their behavioral and cortisol responses, and they showed an increase in habituation over the study period (agitation and cortisol levels both dropped). Individuals in NBH also decreased their responses to observers during focal follows; however, at the end of the study the responses of the two less-habituated groups (NBH and EX) remained elevated in comparison to the habituated group (CP), suggesting the need for further habituation. Unlike capuchin groups that rarely encounter humans, NBH and EX never fled from observers and they rarely emitted observer-directed alarm calls. We suggest that the permanence of habituation and the ability to habituate animals passively through a neutral human presence are both important considerations for researchers conducting studies in areas where animal safety from poachers, etc. cannot be guaranteed.     

Effects of benzocaine and its isomers on sodium permeability and steady state sodium inactivation in the myelinated nerve, obtained by an improved dissection technique

The blocking effects of benzocaine and its isomers (1 mmol/l) on sodium currents in myelinated nerve fibres were tested. As far as the so-called fast sodium inactivation is concerned, benzocaine shifted the h alpha-curve in negative direction to a stronger extent than did its isomers, while the potency of the isomers did not differ significantly from each other. The drug-induced reductions of maximum sodium permeability PNa were tested at constant test pulses at h alpha = 1. In this kind of experiments all the three isomers had the same potencies. The findings could not be correlated to the lipid solubilities of the drugs as measured by the corresponding octanol/water partition coefficients. In addition, efforts were undertaken to minimize any noxious pull during the isolation of the axon. Some consequences of the improvements introduced are discussed in terms of the reliability of ionic current measurements in Ranvier nodes.     

Local anesthetics QX 572 and benzocaine act at separate sites on the batrachotoxin-activated sodium channel

We have studied the effect of local anesthetics QX 572, which is permanently charged, and benzocaine, which is neutral, on batrachotoxin-activated sodium channels in mouse neuroblastoma N18 cells. The dose-response curves for each drug suggest that QX 752 and benzocaine each act on a single class of binding sites. The dissociation constants are 3.15 X 10(-5) M for QX 572 and 2.65 X 10(-4) M for benzocaine. Equilibrium and kinetic experiments indicate that both drugs are competitive inhibitors of batrachotoxin. When benzocaine and QX 572 are present with batrachotoxin, they are much more effective at inhibiting Na+ flux than would be predicted by a one-site model. Our results indicate that QX 572 and benzocaine bind to separate sites, each of which interacts competitively with batrachotoxin.     

Local anesthetics-induced inhibition of chloroplast electron transport

The effects of local anesthetics on photosynthetic activity of pea chloroplasts were investigated in order to elucidate the role of Ca2+ in photosynthetic electron transport. Dibucaine, benzocaine and tetracaine were found to inhibit the O2-evolving activity. The inhibitory effect decreases in the order dibucaine greater than benzocaine greater than tetracaine greater than trimecaine similarly as does the potency to inhibit propagation of excitation in nerve fibre. As demonstrated in experiments with artificial donors and acceptors, the site of inhibition is the water-splitting site of PSII. The inhibitory power of the anesthetics grows with increasing ionic strength of the incubating mixture (by adding NaCl or MgCl2) and with pH; this is explained by occurrence of the neutral form of amine. At low concentrations the charged anesthetic acts as a protonofore; however, the inactivation of water splitting is not due to the protonophoric effect. The incubation is followed by the disappearance of ESR signal IIs. The role of Ca2+ and Ca2+-binding protein in PSII electron transport and its localization are discussed.     

A comparison of the effect of alternating current electronarcosis,rectified current electronarcosis and chemical anaesthesia on the blood physiology of the freshwater bream Oreochromis mossambicus (peters)—III. The effect on the plasma electrolytes Ca2+, Na+ and K+ and on the osmotic pressure of the blood

• 1.1. The effects of alternating current electronarcosis, rectified current electronarcosis and chemical anaesthesia (benzocaine hydrochloride) on plasma electrolytes and on the osmotic pressure of the blood of the freshwater bream Oreochromis mossambicus were evaluated.
• 2.2. Plasma Ca²⁺, Na⁺ and K⁺ concentrations and the osmotic pressure of the blood were monitored over a period of 7 days.
• 3.3. The results showed that the different electrolytes respond differently to the different techniques.
• 4.4. Chemical anaesthesia exhibited the least effects on the parameters studied.

     

血塞通对糖尿病视网膜病变患者视网膜微循环的影响

目的:探究血塞通对糖尿病视网膜病变(DR)患者视网膜微循环的影响。方法:收集2013年2月至2016年2月在我院接受治疗的95例双眼糖尿病视网膜病变患者,并随机分为对照组(48例)和观察组(47例)。对照组患者给予常规降糖治疗,观察组在对照组的基础上给予血塞通治疗。观察比较两组治疗前后的视网膜微循环指标,包括视网膜中央动脉(CRA)的舒张末期血流速度(EDV)、收缩期峰值流速(PSV)及阻力指数(RI)以及视网膜中央静脉(CRV)的最低血流速度(Vmin)、最高血流速度(Vmax)及平均血流速度(MV),血液流变学指标包括全血低切黏度(NBL)、全血高切黏度(NBH)、红细胞变形指数(DE)、红细胞压积(Hct)、红细胞聚集指数(AE)及血沉(ESR)以及临床疗效。结果:治疗后,观察组的临床总有效率为87.8%,明显高于对照组的61.4%(P0.05),对照组的EDV、Vmax、Vmin均较治疗前明显改善(P0.05),NBL和NBH较治疗前明显降低,且观察组的PSV、EDV明显高于对照组,而MV、RI、Vmax、Vmin均低于对照组(P0.05),NBL、NBH、DE、Hct、AE及ESR均较治疗前明显改善,且相较于对照组改善幅度更显著(P0.05)。结论:血塞通联合常规降糖治疗糖尿病视网膜病变患者的疗效显著,可有效改善患者的视网膜微循环和血液流变学。     

General and local anaesthetics perturb the fusion of phospholipid vesicles

The effects of general and local anaesthetics on Ca2+-induced fusion of negatively charged lipid vesicles have been investigated. Vesicles composed of phosphatidylcholine and phosphatidic acid (2:1 molar ratio) were induced to fuse using 5 mM free Ca2+. Fusion, assessed by an increase in size using gel filtration techniques and confirmed by electron microscopy, displayed a dependence on Ca2+ and Mg2+ concentration and on temperature. The inhalational anaesthetics halothane, methoxyflurane and diethyl ether enhanced fusion as did the uncharged local anaesthetic benzocaine. In contrast, the charged local anaesthetics lignocaine and bupivacaine inhibited the fusion process. It is suggested that the enhancement observed with the inhalational anaesthetics and benzocaine was mediated by an effect on lipid fluidity and the inhibition observed with the charged tertiary amine anaesthetics was due to an antagonism towards Ca2+.     

pH-dependent effects of local anaesthetics in perturbing lipid membranes

The effects of local anaesthetics lidocaine, benzocaine, carbisocaine and carbisocaine derivatives, KaQ-7 and Ka-O, in perturbing bovine brain lipid membranes or egg lecithin membranes were compared at pH 6.0; 7.0; and 8.0. The electron spin resonance method with stearic acid labeled at carbon position 16 as the spin probe was employed. The perturbation effects of lidocaine and Ka-O were found to increase with increasing pH of the sample, whereas the effect of carbisocaine decreased with increasing pH. The perturbation effects of benzocaine and KaQ-7 were independent of pH. The pH-dependent perturbation effects of the local anaesthetics tested on lipid membrane fairly corresponded with their pH-dependent potency to block nerve action potentials.