Micro- and Nanomechanical Analysis of Articular Cartilage by Indentation-Type Atomic Force Microscopy: Validation with a Gel-Microfiber Composite

As documented previously, articular cartilage exhibits a scale-dependent dynamic stiffness when probed by indentation-type atomic force microscopy (IT-AFM). In this study, a micrometer-size spherical tip revealed an unimodal stiffness distribution (which we refer to as microstiffness), whereas probing articular cartilage with a nanometer-size pyramidal tip resulted in a bimodal nanostiffness distribution. We concluded that indentation of the cartilage's soft proteoglycan (PG) gel gave rise to the lower nanostiffness peak, whereas deformation of its collagen fibrils yielded the higher nanostiffness peak. To test our hypothesis, we produced a gel-microfiber composite consisting of a chondroitin sulfate-containing agarose gel and a fibrillar poly(ethylene glycol)-terephthalate/poly(butylene)-terephthalate block copolymer. In striking analogy to articular cartilage, the microstiffness distribution of the synthetic composite was unimodal, whereas its nanostiffness exhibited a bimodal distribution. Also, similar to the case with cartilage, addition of the negatively charged chondroitin sulfate rendered the gel-microfiber composite's water content responsive to salt. When the ionic strength of the surrounding buffer solution increased from 0.15 to 2 M NaCl, the cartilage's microstiffness increased by 21%, whereas that of the synthetic biomaterial went up by 31%. When the nanostiffness was measured after the ionic strength was raised by the same amount, the cartilage's lower peak increased by 28%, whereas that of the synthetic biomaterial went up by 34%. Of interest, the higher peak values remained unchanged for both materials. Taken together, these results demonstrate that the nanoscale lower peak is a measure of the soft PG gel, and the nanoscale higher peak measures collagen fibril stiffness. In contrast, the micrometer-scale measurements fail to resolve separate stiffness values for the PG and collagen fibril moieties. Therefore, we propose to use nanostiffness as a new biomarker to analyze structure-function relationships in normal, diseased, and engineered cartilage.     

Beyond the Four Corners of the Convention: Expanding the Scope of Jurisdiction of Law of the Sea Tribunals

In several recent Awards, Part XV tribunals constituted under the UN Convention on the Law of the Sea have been called upon to decide claims based on rights and obligations that find their source in a treaty, agreement, or otherwise binding instrument other than the Convention itself, or on customary international law. This article considers the extent to which such claims fall properly within the jurisdiction of a Part XV tribunal, either on the basis of the applicable law provision in Article 293(1) of the Convention, or on the basis of provisions of the Convention that make reference to other relevant rights and obligations.     

Dissymmetry of the origins of the roots of the spinal cord in the dog

The spinal cord in 25 non-inbred dogs has been studied macro-microscopically. The dissymmetry in the arrangement level in the right and left root bases on the dorsal surface of the spinal cord is much greater than on the ventral surface. The same as in the human being, the dissymmetry is the greatest in the thoracic part (as compared to other spinal parts). On the ventral surface of the spinal cord both along the anterior and posterior margin of the root bases, there is a right-sided dissymmetry (with cranial shift); on the dorsal surface it is present only at the roots along the posterior margin. The dissymmetry of the dog spinal cord is quantitatively estimated along its whole extension.     

Adrenergic and cholinergic components of the innervation of the arteries of the base of the bird brain

By means of Falck's and Koelle's methods adrenergic and cholinergic structures were studied in the arteries in the cerebral basis of blue rock pigeons and of hens, white leghorn stock. The number of nerve transmitters was estimated per 1 mm2 of the vessel surface. The arteries of the basis in pigeon brain are surrounded with developed adrenergic and cholinergic nerve plexuses, their density decreasing in the following order: nasal branch of the internal carotid artery, middle, nasal cerebral and basilar arteries. A little more cholinergic transmitters occur on the middle cerebral artery, while on the other vessels, concentration of cholinergic and adrenergic fibers is equal. In hens, the density in the arrangement of adrenergic nerve transmitters is higher in the nasal branch of the internal carotid and in the nasal cerebral arteries than in the basilar artery. At the same time, chromaffin cells forming numerous conglomerations in some places are found on the latter. In pigeons, the density of adrenergic fibers arrangement on the arteries of the cerebral basis is higher than in hens.     

Characteristics of the membranes of the pellicle of the ciliatePseudomicrothorax dubius

Summary The membranes of the pellicle of the ciliatePseudomicrothorax dubius are investigated using thin section electron microscopy and freeze-fracture replicas. The plasma membrane is covered by a surface coat and is connected to the outer alveolar membrane by short, sometimes branched, bridges. The inner alveolar membrane is coated on both sides. The epiplasm lies in intimate contact with the cytoplasmic surface of this membrane, and there is a corresponding deposit on the other surface. This deposit is regularly striated.The epiplasmic layer and the alveoli are interrupted at sites of cytotic activity,e.g., the attachment sites of trichocysts, the cytoproct, and the parasomal sacs. The striated deposit ends where the epiplasm ends, indicating a direct relationship between these two epimembranous layers.There is a deposit along the sides of the first part of the tip of the trichocysts, and in this region the trichocyst membrane is free of intramembranous particles.The membrane of the parasomal sacs has a coat on both surfaces. That on the extraplasmic surface is similar to the surface coat of the plasma membrane. The origin of the cytoplasmic coat is unknown. The cytotic activity of these sacs is indicated by their highly irregular profiles.     

Functioning of the electromechanical connection in the course of the contracture contraction

The effects of calcium release blocker dantrolene was tested on electrically evoked twitches and on contractures induced by potassium depolarization, by acetylcholine or caffeine. It was shown that the first: developmental, stage of potassium or acetylcholine contracture is inhibited by dantrolene and is not influenced by calcium free medium, therefore we may interpret it as based on a "voltage-dependent Ca release" (VDCR) mechanism of activation, whereas depolarization directly opens the rhyanodin receptor calcium channels. On the contrary, the next stage: the long-lasting plateau of contracture, is directly dependent on external Ca2+ and inhibited by dantrolene, and therefore can be described as "calcium induced Ca-release" (CICR) activation mechanism. In this case stored calcium is also released by rhyanodine receptors, although by means of entering the extracellular Ca2+. Finally, the last stage of low amplitude is not influenced by dantrolene nor by calcium-free medium. Therefore the activation of contraction on this stage is not based on the Ca2+ release through the rhyanodin receptor calcium channels.     

Immunohistochemical localization of the transferrin receptor in the seminiferous epithelium of the rat

The transferrin receptor has been immunohistochemically localized in the seminiferous epithelium of the rat with a monoclonal antibody, MRC OX26, which recognizes the transferrin receptor glycoprotein. The receptor was detectable on mitotically and meiotically dividing germ cells and, less abundantly, on round spermatids. It was lost from germ cells during spermatid elongation and was undetectable on immature spermatozoa. The transferrin receptor was also present on Sertoli cells in the testes of immature animals and on Sertoli cells in the testes of aspermatogenic animals that had been irradiated in utero. It was not detectable on Sertoli cells in the testes of cryptorchid animals. These studies demonstrate that the transferrin receptor is abundant on dividing germ cells as well as dividing somatic cells.     

On the question of cyclic GMP as the mediator of the effects of acetylcholine on the heart

The effects of acetylcholine and sodium nitroprusside on cyclic GMP levels, contractile force, and glycogen metabolism were investigated in the perfused rat heart. While both agents produced time- and concentration-dependent increases in cyclic GMP, only acetylcholine significantly decreased contractile force. Neither agent altered the basal cyclic AMP concentration, cyclic AMP-dependent protein kinase activity ratio, or phosphorylase activity. When dosages were adjusted to give approximately equal increases in cyclic GMP, acetylcholine attenuated the effect of epinephrine on contractile force and glycogen phosphorylase activity while nitroprusside did not antagonize the action of the beta-adrenergic agent on either parameter. The data suggest that increased cardiac cyclic GMP is not sufficient to completely explain the action of acetylcholine on either contractile force or its antagonism of epinephrine-induced increases in force or glycogen phosphorylase activity.     

Autoradiographic study of the ontogeny of the imipramine receptors in the cerebral cortex of rats

Localization of 3H-imipramine binding sites in the cerebral cortex of rats was studied on the 19th day of prenatal development and on the 3d and 14th days of postnatal ontogenesis. Imipramine was injected into rats on days 17, 18 and 19 of gestation. The maximal imipramine receptor density was found in cortex layers IV-V. The therapeutic doses of imipramine provoked marked stimulation of the formation of imipramine receptors on the 3d day of postnatal development. However, on the 14th day the 3H-imipramine binding level was stabilized.     

Effects of the removal of the orbito-frontal cortex on the development of reflex analgesia

The authors studied the effect of electric acupuncture stimulation (EAP) on the changes in pain thresholds prior to and after removal of the orbito-frontal cortex (OFC) of the brain in behavioral experiments on adult cats. Removal of OFC increased the thresholds of pain response at the 4th and the 5th levels of the conventional scale, reflecting emotionally-affective manifestations of pain, and intensified the effect of antinociceptive EAP. The results obtained are analysed in relation to the inhibitory tonic effect of OFC on antinociceptive structures of the brain. Different effects of OFC and somatosensory cortex on the antinociceptive structures of the brain are discussed.     

On the question of cyclic GMP as the mediator of the effects of acetylcholine on the heart.

The effects of acetylcholine and sodium nitroprusside on cyclic GMP levels, contractile force, and glycogen metabolism were investigated in the perfused rat heart. While both agents produced time- and concentration-dependent increases in cyclic GMP, only acetylcholine significantly decreased contractile force. Neither agent altered the basal cyclic AMP concentration, cyclic AMP-dependent protein kinase activity ratio, or phosphorylase activity. When dosages were adjusted to give approximately equal increases in cyclic GMP, acetylcholine attenuated the effect of epinephrine on contractile force and glycogen phosphorylase activity while nitroprusside did not antagonize the action of the beta-adrenergic agent on either parameter. The data suggest that increased cardiac cyclic GMP is not sufficient to completely explain the action of acetylcholine on either contractile force or its antagonism of epinephrine-induced increases in force or glycogen phosphorylase activity.     

Effects of excitation of the central nuclei of the cerebellum on the activity of neurons in the pyramidal tract

The effect of excitation of the contralateral nuclei of the cerebellum on the activity of anti-dromically-identified fast and slow pyramidal neurons in the sensorimotor region (SMR) of the cerebral cortex was investigated in delicate experiments on unnarcotized cats immobilized with procurane. It was found that excitation of the above nuclei could evoke peak responses in the neurons of the pyramidal tract (PT), but more often it modified their background activity. These responses were more pronounced on excitation of the intermediate nucleus (IN) or the dentate nucleus (DN), although they also often arose on excitation of the fastigial nucleus (FN). Qualitatively different effects (stimulative and inhibitory) were observed in some neurons on excitation of different nuclei in the cerebellum. It was found that the nature of the responses by fast neurons in the PT to excitation of the central nuclei in the cerebellum did not differe substantially from the nature of the responses by slow neurons, but that the responses of the latter arose with a longer latent period, and in most cases were less pronounced.L. A. Orbeli Institute of Physiology, Academy of Sciences of the Armenian SSR, Erevan. Translated from Neirofiziologiya, Vol. 3, No. 1, pp. 22–31, January–February, 1971.     

Effect of the chemical nature of the carbon source in the medium on the makeup of the biomass]

The yeast Candida tropicalis, Candida utilis, Trichosporon cutaneum were cultivated on the synthetic medium containing additions of group B vitamins (biotin, thiamin) and glucose, glycerol of lactic acid as the sole source. The protein content, amino acid composition, nucleic acids and complex B vitamins were identified in the resultant biomass. The carbon source in the medium affected these indices. The protein and nucleic acid content also depended on the yeast strain.     

Changes in the digestibility of constituents of the diet in the rabbit]

Fiber digestibility of two diets (the first based on alfalfa dried, the second based on beet-pulp dried) with the same level of crude fiber and crude protein was measured on rabbits of 6, 7, 11, 13 and 17 weeks old. The age has no significative effect on digestibility (dry matter, organic matter, crude fiber) of the two experimental diets. The quantity of cellulose and hemicellulose digested was greater in rabbits on the pulp diet. Independantly of the diet hemicellulose is more digestible than cellulose. Expediency of use other analytical test than crude fiber in the nutritional requirements of rabbits is discussed.     

The participation of the GABA-A and GABA-B receptors in the mechanism of the inhibition of the contractile activity in the rabbit myometrium under the influence of GABA, AOAA and fenibut

Experiments on isolated strips of the rabbit uterus showed the stimulating effects of small doses of GABA, AOAA and phenibut on uterine contractility, while large doses exerted the reverse (suppressing) effects. Administration of bicuculline and picrotoxin before or after the above-mentioned drugs reduced their suppressing effects on uterine muscle contractility. The data postulate the involvement of GABAA and GABAB receptors in the drugs action on the rabbit uterus.     

Influence of the redox potential of the medium on the activity of polyphenol oxidase

While ascorbate is an inhibitor of polyphenol oxidase upon its activity on tyrosine, hydrogen peroxide has a promoting effect especially in conjunction with catalase. When both ascorbate and peroxide are present, the effect of the former dominates over the latter. These influences can be explained on the basis of the redox potential of the solutions: high potentials promote, while low potentials decrease the rate of the oxidative enzyme reaction. The findings might have some implications on the process of aging of tissues.     

试论生物经济成长(GREW)战略

生命科学与生物技术的发展推动了"生物经济"概念的形成与生物经济时代的来临,进而导致农业、健康医疗、环保、工业制造等产业的生产与消费方式正在发生深刻变革。分别从生物经济对未来农业、健康医疗、环保及制造业的影响进行分析,提出了生物经济成长(GREW)战略。作为一个战略体系,GREW战略包括相互交叉、相互依存与影响的四个子战略:新型农业(Green biotech based)战略、健康医疗(Red biotech based)战略、绿色环保(Grey or Environmental biotech based)战略,以及绿色制造业(White biotech based)战略。GREW战略的及时提出与逐步形成,对于生物产业发展规划及政策制定具有高度前瞻性意义和指导作用。     

Intraoperative control of the state of the arrested heart

Monitoring the function of the operated on heart over time based on the principle of recording the effort of the myocardium at rest was tried under the clinical conditions after preliminary experiments on the hearts of rats and dogs. The study included 11 patients operated on for aortal, mitral and multi-valvular prosthetics under pharmacological cardioplegia. Such monitoring enables an objective appraisal of the heart status, Besides, it makes it possible to decide on whether repeated administration of cardioplegic solutions is required, to specify the times of reoxygenation and to prognose the pattern of the heart function recovery after operation is over. The data obtained helped detect some disadvantages of the generally accepted method of pharmacological cardioplegia.     

A study of the regulation of photosynthesis by the analysis of the effect of kinetic parameters on the characteristics of the oscillatory regime and the rate of CO2 assimilation

Earlier at the biophysics department, the experimental data on the oscillations of delayed luminescence have been described with the help of a mathematical model. Here we studied the influence of the model parameters on the characteristics of the oscillatory regime. The frequencies and damping factors of the oscillations at different parameter values were calculated using the Lyapunov analysis. It was shown that, in addition to oscillations observed experimentally, other, rapidly damping oscillations may exist. The dependence of the CO2 assimilation rate on the model parameters was studied. It was shown that the intensity of the light absorbed by photosystems I and II may differently affect the assimilation of CO2.     

Properties of the on-transient of the intracellular response in the barnacle photoreceptor

We have studied the on-transient of the receptor potential of the barnacle photoreceptor. Its amplitude has previously been shown to depend on light intensity and state of light-dark adaptation. We have examined its dependence on 1) the presence of a prolonged depolarizing afterpotential (PDA), 2) a background light, 3) added alcohol, or 4) decreased K⁺ concentration in the bath. We find that the relative on-transient amplitude tends to increase initially with increasing depolarization arising from 1)–4) and then to decrease again at higher depolarization. This behavior is qualitatively explainable by the cell's currentvoltage characteristics and by the adapting effect of the stimulus on the conductances arising from the PDA, the background light and the alcohol.Based on material presented at the European Neurosciences Meeting, Florence, September 1978