Identification of chemosensory sensilla mediating antennular flicking behavior in Panulirus argus, the Caribbean spiny lobster

Crustaceans sample odorants by a rapid series of flicks of the two flagella composing the distal segments of each of the paired antennules. The lateral flagella contain aesthetasc sensilla that house unimodal chemosensory neurons. Nine types of nonaesthetasc setae with putative chemosensory and mechanosensory functions are distributed on the lateral and medial flagella. Sensory neurons in aesthetascs and nonaesthetasc sensilla terminate in separate regions of the brain, the olfactory lobe, and the lateral antennular neuropil, resulting in two odorant-processing pathways. Distilled water ablation of flagella and excision of specific setae were used to identify chemosensory sensilla mediating antennular flick behavior in Panulirus argus. The flick rates of sham-ablated and ablated or excised lobsters toward squid extract were compared. Complete attenuation of flick response to squid extract occurred as a result of (1) distilled water ablation of lateral flagella, (2) excision of aesthetascs and asymmetric sensilla, and (3) excision of aesthetascs. Distilled water ablation of medial flagella resulted in a mean flick rate 52% of that observed for sham-ablated lobsters toward squid extract. Flicking was unaffected by excision of asymmetric, guard, or companion sensilla. We propose that odorant mediation of flicking behavior requires both the aesthetasc and nonaesthetasc pathways.     

Isolation of a factor stimulatory to Bradyrhizobium japonicum in broth culture

Previous experiments indicated that water extracts of lambsquarters (Chenopodium album) and green foxtail (Setaria viridis), among others, stimulated growth of Bradyrhizobium japonicum in broth culture. Water extracts of lambsquarters shoots collected before or after anthesis were equally stimulatory. The stimulatory effect of extracts of lambsquarters when heated to 100°C for 30 min or autoclaved for 15 min was reduced by about 20% compared to untreated extracts. Extracts of green foxtail were less affected by higher temperature under similar conditions. Extracts of both green foxtail and lambsquarters completely lost their stimulatory effect following exposure to aerial microflora for 120 h. Water extract of lambsquarters shoots was more stimulatory than methanol extract, and neither ether nor butanol extracts resulted in stimulation. Both shoots and roots of lambsquarters and green foxtail were sequentially extracted first by water followed by methanol and vice-versa. The bioassay of these extracts indicated that there could be two components of the growth factor-one, larger component is soluble in water, the other, smaller component is soluble in methanol. After fractionation of the crude aqueous extract of lambsquarters shoots by four organic solvents, the residual aqueous extract retained the growth factor. Dialysis of the residual aqueous extract of lambsquarters shoots through a membrane (MWCO 1000) indicated that the molecular weight of the growth factor was less than 1000. The fraction having molecular weight <1000 was separated by paper chromatography using 6% acetic acid as developer. The fraction with Rf 0.91 showed the highest stimulation of the bacterium.     

Innervation pattern of the deep extensor abdominal muscle in the crayfish species Astacus astacus

The deep extensor abdominal muscle consisting of one medial and two lateral muscle bundles together with the nerve innervating the muscles of crayfish species Astacus astacus, was prepared. Light microscopic investigations of methylene blue stained preparations showed that the nerve innervating the deep extensor abdominal muscle consists of five distinct axons. The five axons were stained separately with lucifer yellow and the innervation pattern of the axons was determined. To confirm the histological results the axons were also stimulated with a suction electrode to elicit excitatory postsynaptic currents on the muscle membrane which were detected using a macro patch electrode. The muscle is innervated by a common excitatory and a common inhibitory axon branching over all three muscle bundles and sending additionally a branch to the L1-bundle of the next posterior segment, and by two axons specific for the two lateral muscle bundles. The axon specific for the innervation of the L1-bundle sends also a branch to the L1-bundle of the next posterior segment. In addition there is one excitatory axon which directly innervates the medial muscle bundle of the next posterior segment branching in most of the cases also to the medial bundle of the segment where it originates.Abbreviations DEAM deep extensor abdominal muscle - EPSC excitatory postsynaptic current - IPSC inhibitory postsynaptic current - L lateral - M medial - GABA -aminobutyric acid     

Role of olfaction and vision cues in feeding behavior and alarm reaction in the catfish pintado, Pseudoplatystoma corruscans

Experiments were performed to investigate senses that are essential for mediating fright reaction and food behavior in Pseudoplatystoma corruscans, pintado. The dilemma “to feed or to flee” was also analyzed in fishes with intact and sectioned olfactory tracts, stimulated by alarm substance extracts and food. Fishes were arranged into five groups: fish with intact lateral olfactory tracts (LOT), fish with intact medial olfactory tract (MOT), fish with tracts totally sectioned (TOTAL, both LOT and MOT), sham operated, and nonoperated fish. The five groups were submitted to either alarm substance extract and food stimulus or to distilled water (control) and food stimulus. Fish reacted to food independently of which tract (LOT, MOT or TOTAL) was sectioned; vision seems necessary and elemental to detect and deflagrate food response. Latency of the responses to each reaction was different between groups. None of the fish with sectioned tracts reacted to alarm substance extract, while sham- and nonoperated fish showed the typical alarm behavior response, leading to the conclusion that olfaction is essential for mediating alarm response. These results indicate that others sense systems (e.g., vision) are sufficient to trigger and elicit feeding behavior and that olfaction is not necessary to fully maintain food detection to qualitative and quantitative extent. However, olfactory tract integrity seems to be required for mediation of alarm reaction in P. corruscans.     

Neurotrophic effect of nerve extract on development of tetrodotoxin-sensitive spike potential in skeletal muscle cells in culture.

The effect of the presence of nerve extracts on the development of tetrodotoxin (TTX)-sensitive sodium channels in cultures of dissociated embryonic chick skeletal muscle cells was examined by measuring the maximum rate of rise of TTX-sensitive spike potential. The addition of the nerve extract prepared from brain or spinal cord of chick embryos to the culture medium caused an increase in the channel density. Extracts of non-neural tissues, i.e., lung, kidney, and muscle, were ineffective. Liver extract, however, produced an effect similar to the nerve extracts. These results suggest that the TTX-sensitive sodium channels in the muscle cell membrane are regulated by a diffusible chemical substance independently of innervation, and that this substance resides in neural tissues, and perhaps also in liver.     

Innervation and motor patterns of the abdominal superficial flexor muscles in larval lobsters

The pattern of innervation and motor program of the abdominal superficial flexor muscle was investigated electrophysiologically in larval lobsters (Homarus americanus). The muscle receives both excitatory and inhibitory innervation in the larval as well as in the embryonic stages. Individual muscle fibers receive a single inhibitory neuron (f5) and a maximum of three excitors. Based on spike heights these axons belong to either the small (f1 or f2) or large (f3, f4) motoneurons. While the small axons preferentially innervate the medial muscle fibers the large axons innervate medial as well as lateral fibers. This larval pattern of innervation resembles the pattern in the adult lobster. The resemblance extends to the firing patterns as well with both large and small excitors firing spontaneously. Furthermore, evoked activity in the larvae produces reciprocal (and occasionally cyclical) bursts of excitor and inhibitor neurons denoting abdominal extension and flexion and resembling the firing patterns in adults. Consequently motor programs employed in steering the pelagic larvae are reminiscent of the programs for maintaining posture in the benthic adult lobsters.     

富贵竹提取液中几种成分的初步研究

在富贵竹枝条基部提取液中，ＩＡＡ含量和酚类物质含量显著地高于中部和上部提取液的含量；中部提取液中的可溶性糖含量和蛋白质含量最高．     

Topography and nerve supply of the cucullaris (trapezius) of skates

Dissections of Sudan black B stained specimens reveal that, of a complex of medial, intermediate, and lateral muscles of skates, presumed homologous to the cucullaris of sharks, only the lateral muscle is innervated by a branch or branches of the vagus and is inserted, in part, to the fused pharyngobranchials of the caudal visceral arches. The medial and intermediate muscles are supplied by separate branches of rostral spinal nerves and do not attach to the branchial skeleton. The lateral muscle therefore is the most likely homologue of the cucullaris (trapezius) of sharks and perhaps other fishes and tetrapods. The medial and intermediate muscles appear to be part of the axial musculature.     

The first branch of the lateral plantar nerve and heel pain

The course and ramification pattern of the lateral plantar nerve was studied in serial sections from 4 fetal feet and in dissections from 34 adult feet with special reference to the so called first branch. This branch was found in all of the observed fetal and adult specimen. From its originating point the nerve runs immediately distally to the medial process of the calcaneal tuberosity in a lateral direction to the proximal part of the abductor digiti minimi muscle. During its course the FB gives two branches. One of them penetrates sometimes the insertion of the quadratus plantae muscle, whereas in adult feet it always sends fibres to the periosteum around the medial process of the calcaneal tuberosity and the long plantar ligament. The other innervates the flexor digitorum brevis muscle. The site of a possible entrapment is located between the abductor hallucis muscle and the medial head of the quadratus plantae muscle. There is strong indirect evidence that the nerve is of a mixed type consisting of sensory fibres for the calcaneal periosteum and the medial head of the quadratus plantae muscle. There is strong indirect evidence that the nerve is of a mixed type consisting of sensory fibres for the calcaneal periosteum and the long plantar ligament as well as motor fibres for the quadratus plantae, flexor digitorum brevis and abductor digiti minimi muscles, which may explain the characteristic pain complaints of the heel pain syndrome. The occurrence of a stiff fascia perforated by the nerve branch or a bursa around the insertion of the plantar aponeurosis as has been described by several authors and which was put forward as a possible aetiological factor could not be confirmed in our material.     

泽泻汤提取物三萜类化学成分分析及其抗非酒精性脂肪肝作用研究

目的:对泽泻汤不同极性溶剂提取物三萜类化学成分进行定性分析并探究其对非酒精性脂肪肝的干预作用。方法:制备泽泻汤水及95%乙醇提取物,利用超高效液相色谱串联高分辨质谱技术对三萜类成分进行定性鉴别,并比较两种提取物中三萜类化合物数量及含量差异。采用油酸诱导的HepG2细胞脂肪累积模型评价泽泻汤不同极性提取物干预非酒精性脂肪肝的能力。结果:在泽泻汤中共鉴定20个三萜类色谱峰,水提取物中鉴定出14个三萜类成分,其中16-oxo-alisol A相对含量最高,95%乙醇提取物中鉴定出16个三萜类成分,alisol B 23-acetate相对含量最高。两提取物均能显著降低细胞中的脂肪累积量,且乙醇提取物表现出更强的抑制细胞内脂肪累积的作用。结论:泽泻汤水及95%乙醇提取物中三萜类化合物数量和含量存在差异,两种提取物均具有潜在抗非酒精性脂肪肝作用,且乙醇提取物活性更强,研究结果为泽泻汤进一步的药效物质基础及药理活性研究提供了基础。     

Volatile substances from adult extracts induce larval settlement of the barnacle balanus amphitrite *

The volatile substance extracted from conspecific adults induces larval settlement of the barnacle Balanus amphitrite. The settlement inducing activity of the volatile fractions was checked monthly from June, 1997 to December, 1998. Both water soluble extracts and volatile fractions from the barnacle were prepared by the steam distillation. The active cue in the volatile fraction was always extracted with n‐pentane under acidic conditions, although settlement inducing activity varied with the sample. GC‐MS analysis of the active and inactive pentane fractions revealed 1, 2, 3‐trimethyl‐benzene as the settlement inducing substance. Commercially available 1, 2, 3‐trimethylbenzene also showed high settlement inducing activity at a concentration of 0.8 × 10^‐12 M (100 pg 1^‐1). This substance was detected at concentrations of more than 7 ng g^‐1 of wet barnacle (equivalent to 0.6 × 10^‐12M, equal to 70pg1^‐1) by GC analysis. These results indicate that 1,2,3‐trimethylbenzene in the volatile fractions acts as a chemical cue for larval settlement. Monthly variation in the settlement inducing activity was observed, which synchronized with the breeding season of the barnacle. This observation suggests that the barnacle produced the chemical cue in the gonad during maturation or accumulated it from the environment.     

Surgical anatomy of the gracilis muscle of the thigh

As demonstrates the investigation of 100 preparations, the main vascular-nervous "hilus" of the muscle is situated on the superior third of its lateral part. They contain the transversal branch of the medial artery, circumflexing the femoral bone with the accompanying vein, having the same name, and the anterior branch of the obturative nerve (92 preparations), or muscle branch of the femoral nerve (8 preparations). The muscle has a well developed network of intrasystemic and intersystemic anastomoses. Beginning from the first period of childhood, for sphincter plasticity either the whole muscle, or a graft, cut out from its lateral part, can be used.     

Involvement of a low-molecular-weight substance in in vitro activation of the molybdoenzyme respiratory nitrate reductase from a chlB mutant of Escherichia coli.

The soluble subcellular fraction of a chlB mutant contains an inactive precursor form of the molybdoenzyme nitrate reductase, which can be activated by the addition to the soluble fraction of protein FA, which is thought to be the active product of the chlB locus. Dialysis or desalting of the chlB soluble fraction leads to the loss of nitrate reductase activation, indicating that some low-molecular-weight material is required for the activation. The protein FA-dependent activation of nitrate reductase can be restored to the desalted chlB soluble fraction by the addition of a clarified extract obtained after heating the chlB soluble fraction at 100 degrees C for 8 min. The heat-stable substance present in this preparation has a molecular weight of approximately 1,000. This substance is distinct from the active molybdenum cofactor since its activity is unimpaired in heat-treated extracts prepared from the organism grown in the presence of tungstate, which leads to loss of cofactor activity. Mutations at the chlA or chlE locus, which are required for molybdenum cofactor biosynthesis, similarly do not affect the activity of the heat-treated extract in the in vitro activation process. Moreover, the active material can be separated from the molybdenum cofactor activity by gel filtration. None of the other known pleiotropic chlorate resistance loci (chlD, chlG) are required for the expression of its activity. Magnesium ATP appears to have a role in the formation of the active substance. We conclude that a low-molecular-weight substance, distinct from the active molybdenum cofactor, is required to bestow activity on the molybdoenzyme nitrate reductase during its biosynthesis.     

Differential activation of parts of the latissimus dorsi with various isometric shoulder exercises

As no study has examined whether the branches of the latissimus dorsi are activated differently in different exercises, we investigated intramuscular differences of components of the latissimus dorsi during various shoulder isometric exercises. Seventeen male subjects performed four isometric exercises: shoulder extension, adduction, internal rotation, and shoulder depression. Surface electromyography (sEMG) was used to collect data from the medial and lateral components of the latissimus dorsi during the isometric exercises. Two-way repeated analysis of variance with two within-subject factors (exercise condition and muscle branch) was used to determine the significance of differences between the branches, and which branch was activated more with the exercise variation. The root mean squared sEMG values for the muscles were normalized using the modified isolation equation (%Isolation) and maximum voluntary isometric contraction (%MVIC). Neither the %MVIC nor %Isolation data differed significantly between muscle branches, while there was a significant difference with exercise. %MVIC was significantly higher with shoulder extension, compared to the other isometric exercises. There was a significant correlation between exercise condition and muscle branch in the %Isolation data. Shoulder extension and adduction and internal rotation increased %Isolation of the medial latissimus dorsi more than shoulder depression. Shoulder depression had the highest value of %Isolation of the lateral latissimus dorsi compared to the other isometric exercises. Comparing the medial and lateral latissimus dorsi, the medial component was predominantly activated with shoulder extension, adduction, and internal rotation, and the lateral component with shoulder depression. Shoulder extension is effective for activating the latissimus dorsi regardless of the intramuscular branch.     

EVALUATION OF PIGMENT EXTRACTION METHODS AND A RECOMMENDED PROTOCOL FOR PERIPHYTON CHLOROPHYLL a DETERMINATION AND CHEMOTAXONOMIC ASSESSMENT1

Many methods have been proposed to extract and quantify algal pigments. Comparative studies have found that pigment extraction efficiency varies among solvent and mechanical disruption protocols due to differential cellular resistance, thereby, leading to potential misinterpretation of pigment data. When the type or resistance of algae are unknown, a method is required that efficiently extract pigments from all taxonomic groups. The objective of this study was to develop a simple and efficient one stage periphyton pigment extraction protocol by comparing the extractability of four solvents (acetone, methanol, methanol/acetone, and methanol/acetone/N,N‐dimethylformamide), the effects of grinding, and the effects of freeze‐drying. The best overall extraction was obtained using freeze‐dried samples extracted with methanol/acetone/DMF/water (MAD). Eighty‐six percent more chlorophyll was extracted when the sample was freeze‐dried relative to fresh/frozen samples extracted with 90% acetone. Freeze‐drying greatly improved the extraction of both polar and non‐polar (lipophilic/hydrophobic) pigments while MAD increased the extractability of polar pigments and improved peak resolution of all pigments. Chemotaxonomic assessment differed between samples that were fresh/frozen or freeze‐dried before extraction. The relative abundance of cyanobacteria was greater for freeze‐dried material compared with fresh/frozen due to the improved extractability of cyanobacterial pigments. Based on the results of this study, the traditional approach of 90% acetone as a solvent is not recommended for periphyton samples containing cyanobacteria or when the composition of the mat is unknown. The combination of freeze‐drying and MAD was sufficient for the extraction of pigments from a periphyton mat containing filamentous cyanobacteria, green algae, and diatoms.     

不同植物药提取物对灵芝细胞生长和胞内三萜产物形成的影响

研究了不同植物药的水提和醇提物对灵芝深层发酵过程中菌丝量和胞内三萜产量的影响。将不同植物药的水提物和醇提物分别加入到发酵基础培养基中,培养7d后检测灵芝生物量和胞内三萜含量。结果表明,金银花和枸杞子水提物添加浓度为100mg/L时,可促进灵芝细胞的生长（p＜0.05）。连翘水提物对灵芝生长和胞内三萜的形成都有显著促进作用,当连翘水提物浓度为400mg/L时,胞内三萜产量从对照的（192.54±8.99）mg/L提高到（302.52±3.79）mg/L。金银花和枸杞子醇提物浓度为200mg/L时能显著促进灵芝细胞生长;枸杞子醇提物在同样浓度下还能促进灵芝胞内三萜的形成。但板蓝根和银杏叶水提物和醇提物都对灵芝的细胞生长和胞内三萜形成有较强的抑制作用。     

Projections and actions of tachykininergic, cholinergic, and serotonergic neurones in the intestine of the Atlantic cod

The native tachykinins cod neurokinin A and cod substance P, serotonin and acetylcholine have excitatory effects on the circular smooth muscle of the cod intestine. Furthermore, immunoreactivities to the cod tachykinins, serotonin and two markers for cholinergic neurones, viz. choline acetyltransferase and vesicular acetylcholine transporter, have been demonstrated in myenteric neurones of the cod intestine. In order to elucidate whether the neurones containing these substances project orally and thus might be involved in the ascending excitatory reflex of peristalsis, myotomy operations have been performed on the cod intestine. The immunoreactive areas of the myenteric plexus immediately oral and anal to the myotomy operations have been measured by using confocal laser scanning microscopy. Large accumulations of immunoreactivity to the tachykinins are found on the anal side of the myotomies, indicating oral projections of tachykininergic neurones. The areas immunoreactive to serotonin and choline acetyltransferase are of equal size on the oral and anal sides. Since the tachykinin containing neurones of the intestine project orally, and since cod neurokinin A and cod substance P have excitatory effects on circular smooth muscle, we conclude that tachykininergic neurones are involved in the ascending excitatory reflex of peristalsis in the cod intestine. Received: 6 March 1997 / Accepted: 15 September 1997     

几种昆虫水提和醇提物对灵芝深层发酵生产多糖的影响

研究了几种昆虫的水提物和醇提物对灵芝深层发酵生产菌体和多糖的影响。结果表明,松毛虫水提物在剂量为30mg/L时,能促进灵芝菌体的生长,生物量从15.23±0.53g/L提高到16.26±0.67g/L。其它昆虫提取物对灵芝菌体的生长无显著影响,其中斑蝥提取物表现出明显的抑制作用。对灵芝多糖产量的影响试验表明,蜣螂水提物在剂量为50mg/L时,能显著提高灵芝胞内多糖的产量,产值从1.93±0.09g/L提高到2.34±0.13g/L;地鳖虫醇提物在剂量为55mg/L,以及松毛虫醇提物在剂量为35mg/L时,对胞内多糖的生产也具有促进作用。此外,蜣螂醇提物在剂量为20mg/L时,对胞外多糖的生产具有促进作用,产量从520.3±20.2g/L提高到589.5±24.1mg/L。结果提示药用昆虫中含有促进灵芝多糖生物合成的功能因子。     

Flagellar motility requires the cAMP-dependent phosphorylation of a heat-stable NP-40-soluble 56 kd protein, axokinin

Using NP-40-treated dog sperm as a model, the stimulatory effect of cAMP upon reactivated flagellar motility has been shown to be dependent upon the cAMP-dependent phosphorylation of a heat-stable NP-40-soluble protein of 56 kd. Examination by two-dimensional polyacrylamide gel electrophoresis of NP-40 extract proteins phosphorylated with gamma-32P-ATP revealed a major cAMP-dependent phosphopeptide at 56 kd. This is the only cAMP-dependent phosphoprotein common to NP-40 extracts of all tissues that show cAMP-dependent stimulation of flagellar motility. These cells and tissues include sea urchin, dog, and human sperm, as well as dog trachea and retina. Moreover, this phosphoprotein is absent in nonstimulatory extracts from tissues such as skeletal muscle, brain, and liver. We conclude that the cAMP-dependent phosphorylation of the 56 kd peptide represents a major regulatory component of not only sperm but other types of axonemal motility as well.     

小桐子提取物除草活性的生物测定

为全面了解小桐子(Jatropha curcas L. )提取物的除草活性,以萝卜(Raphanus sativus L. )、苋(Amaranthus tricolor L. )、苏丹草[Sorghum sudanense (Piper) Stapf]和黑麦草(Lolium perenne L. )为实验材料,对小桐子果壳和枝叶的水、乙醇(体积分数95%)、正丁醇、乙酸乙酯、氯仿和石油醚粗提物的除草活性进行了生物测定,并从中筛选出抑制作用最强的水粗提物进行进一步的活性组分分离及其除草活性的生物测定.测定结果显示,小桐子果壳和枝叶的6种溶剂提取物(10 g·L~(-1))对供试的4种植物幼苗的根长和茎高均有不同程度的抑制作用,其中水粗提物和乙醇(体积分数95%)粗提物的抑制作用较强,且水粗提物对供试的4种植物幼苗的根长和茎高的抑制作用均在75%以上,显著高于其他溶剂粗提物(P<0.05);石油醚粗提物的抑制作用最小,均在10%以下.小桐子果壳和枝叶水粗提物的石油醚、氯仿、乙酸乙酯、正丁醇和水萃取物(10 g·L~(-1))对萝卜和苏丹草幼苗的根长和茎高均有不同程度的抑制作用,其中水、正丁醇和乙酸乙酯萃取物的抑制作用显著高于氯仿和石油醚萃取物,抑制率均在70%以上;水萃取物的抑制作用最强,抑制率均在80%以上;石油醚萃取物的抑制作用最小,抑制率均在10%以下.研究结果表明,小桐子果壳和枝叶的水粗提物具有一定的除草活性,其有效成分为极性较大的组分.