ULTRASTRUCTURAL STUDIES ON THE PERMEABILITY OF THE MESOTHELIUM TO HORSERADISH PEROXIDASE

Peritoneal mesothelium was exposed for 2–60 min to solutions of horseradish peroxidase by incubation in vitro, or after intraperitoneal injection in vivo. Peroxidase was localized, with the electron microscope in the intercellular clefts of the mesothelium, often along their entire lengths, in vesicles adjoining or contiguous with the clefts, and along the peritoneal and basal surfaces of the cell, and also in intracytoplasmic vacuoles. The intercellular junctions of peroxidase-treated mesothelium did not differ from those of controls: open and closed junctions were present in both groups. Intercellular localization was also obtained when the mesothelium was exposed to peroxidase during or after fixation. Although intracellular absorption of peroxidase and its incorporation into larger vacuoles were observed, there was no clearcut evidence of vesicular transport across the mesothelium in these experiments. These findings are consistent with physiologic data which postulate that mesothelial transport can be accounted for, at least in part, by passive diffusion through a system of pores, and they suggest that these pores are located in the intercellular clefts.     

THE NUCLEAR ANNULI AS PATHWAYS FOR NUCLEOCYTOPLASMIC EXCHANGES

Colloidal gold particles, 25 to 55 A in diameter, which had been coated with polyvinylpyrrolidone, were microinjected into the ground cytoplasm of amebas (Chaos chaos). At time intervals of 1 minute, 2 minutes, 10 minutes, and 24 hours after injection the cells were fixed for electron microscopy. After 24 hours, gold particles were found in both the nuclei and the ground cytoplasm, the concentration being higher in the nuclei. Colloidal particles were also present in the nuclei after 10 minutes, but at this time interval the concentration did not appear to be greater than that in the ground cytoplasm. One and 2 minutes after injection, the gold particles were located almost exclusively in the ground cytoplasm; however, individual particles were often found within the annuli of the nuclear envelope, and were located specifically in the centers of these structures. The results suggest that at least some of the gold particles which enter the nuclei pass through the annuli, and that passage through these structures may be restricted to a central channel.     

脑室注射纳洛酮对大鼠迷走-加压反应的影响

本文研究了太鼠的迷走-加压反应和脑室注射纳洛酮对大鼠迷走-加压反应的影响。其结果为:1.刺激大鼠迷走神经向中端,可出现迷走-加压反应;2.脑室注射纳洛酮15—20分钟左右,大鼠迷走-加压反应显著抑制;50分钟左右抑制效应解除,迷走-加压反应开始复现。以上事实提示:内源性阿片样物质参与大鼠的迷走-加压反应过程,对迷走-加压反应可能起加强作用。     

IDENTIFICATION OF THE ADRENOCORTICOTROPHIN-PRODUCING CELLS IN THE RAT HYPOPHYSIS BY AUTORADIOGRAPHY

The relative rates of protein (hormone) synthesis and secretion by the various cell types in the anterior hypophysis of the rat have been studied by means of autoradiography. Normal and adrenalectomized male rats were injected with tritiated glycine and their hypophyses removed and fixed at 20, 40, and 90 minutes and 15 hours after injection. Autoradiograms of the hypophysial sections were prepared and autoradiographic grains were counted in the film overlying the cytoplasm of individual cells. With the aid of this method, a unique cell type was identified in the hypophyses of adrenalectomized rats. This cell is morphologically distinct from "gonadectomy cells," "thyroidectomy cells," and from previously described normal cell types, and is therefore designated as the "adrenalectomy cell." Among the 7 cell types differentiated in this study, the "adrenalectomy cell" had the highest tritium content and, in addition, at the time intervals studied this cell had the fastest rate of appearance and disappearance of protein tritium. This autoradiographic evidence of rapid protein (or polypeptide) turnover following adrenalectomy indicates that the "adrenalectomy cell" is the site of adrenocorticotrophin production in the adrenalectomized rat. Further autoradiographic and cytological evidence is presented which suggests that the "adrenalectomy cells" may be derived from chromophobes, and that a portion of the "large chromophobes" as defined in this study may be the site of adrenocorticotrophin production in the normal rat.     

A role of the rostral hypothalamus in the regulation of LHRH release in baboons

In the intact control baboons the plasma level of LH was elevated and reached a peak within 15 minutes after administration of 100 microgram synthetic LHRH. In addition, a second peak in plasma LH appeared within 90 minutes after LHRH injection. Supplementing these results, plasma level of estrogen was elevated within 30 minutes after LHRH injection. Subsequently, frontal deafferentation of the hypothalamus was performed in these baboons. After administration of 100 microgram synthetic LHRH in these frontally deafferented baboons the plasma level of LH was elevated and reached a peak within 15 minutes, as in the intact control baboons. However, there was no second LH peak within 90 minutes after LHRH injection, even though plasma estrogen was elevated within 30 minutes, as in the intact control baboons. It was found that the rostral hypothalamus in baboons is involved in the regulation of LHRH release which promotes to release LH within 90 minutes after LHRH injection.     

~(125)Ⅰ—蜂毒肽在小鼠体内分布、吸收、排泄的研究

以氯胺T为氧化剂参照Hunter和Greenwood的多肽类化合物的碘化标记法,制备了~(125)I—标记的蜂毒肽在小鼠体内分布、吸收、排泄的研究,实验证明小鼠肌注~(125)I—蜂毒肽后,吸收很快,主要分布部位为肾、肺、心、肝、小肠、关节、脾与肌肉,脑组织中含量很少,肌肉注射后5分钟血液中含量可达70％,~(125)—I蜂毒肽主要经肾排泄,肌注后30分钟肾脏浓集最高,而尿液中以1.5小时为最高,而粪便中排泄少。     

OBSERVATIONS ON THE UPTAKE OF TRITIATED THYMIDINE IN THE PRONUCLEI OF FERTILIZED SAND DOLLAR EMBRYOS

Following fertilization of the egg of the sand dollar Echinarachnius parma, tritiated thymidine (H³TDR) was taken up independently by the male and female pronuclei beginning within about 15 to 20 minutes, and the labeled pronuclei fused at about 30 to 40 minutes. At cleavage 90 minutes later the labeled nuclear material was distributed to both daughter cells. Unfertilized eggs and sperm exposed to H³TDR did not show nuclear localization of thymidine. DNA replication, thus, is initiated in the haploid pronuclei shortly after fertilization and prior to fusion. The major portion of DNA synthesis, as evidenced by thymidine uptake, appears to be during a 20 to 30 minute period after fertilization. Fertilization is associated with the activation of a mechanism which initiates early and independent replication of DNA in both the male and female pronuclei.     

THE FINE STRUCTURE OF THE AXON AND GROWTH CONE OF THE DORSAL ROOT NEUROBLAST OF THE RABBIT EMBRYO

The centrally directed neurite of the dorsal root neuroblast has been described from the period of its initial entrance into the neural tube until a well-defined dorsal root is formed. Large numbers of microtubules, channels of agranular reticulum, and clusters of ribosomes are found throughout the length of the early axons. The filopodia of the growth cone appear as long thin processes or as broad flanges of cytoplasm having a finely filamentous matrix material and occasionally small ovoid or elongate vesicles. At first the varicosity is a small expansion of cytoplasm, usually containing channels of agranular reticulum and a few other organelles. The widely dilated cisternae of agranular reticulum frequently found within the growth cone probably correspond to the pinocytotic vacuoles seen in neurites in tissue culture. The varicosities enlarge to form bulbous masses of cytoplasm, which may measure up to 5 µ in width and 13 µ in length. They contain channels of agranular reticulum, microtubules, neurofilaments, mitochondria, heterogeneous dense bodies, and a few clusters of ribosomes. Large ovoid mitochondria having ribonucleoprotein particles in their matrix are common. Dense membrane specializations are found at the basal surface of the neuro-epithelial cell close to the area where the early neurites first enter the neural tube.     

THE DISTRIBUTION WITHIN THE BRAIN OF FERRITIN INJECTED INTO CEREBROSPINAL FLUID COMPARTMENTS : I. Ependymal Distribution

From 10 minutes to 3½ hours after the intraventricular injection into rats of 15 to 100 mg of ferritin, an appreciable fraction of the protein, visualized electron microscopically, traverses the ependymal epithelium by diffusing along the dense intercellular substance of the luminal open junction and thence, by circumventing discrete intercellular fusions which partition rather than seal the interspace. These partitions shunt additional protein into the cell, where ferritin is transported within pinocytotic vesicles to the lateral and basal plasma-lemma and, presumably, back into the interspace again. The basal interspace is irregularly distended by pools of moderately dense "filler" within which ferritin accumulates. The larger fraction of protein enters the ependyma by pinocytosis and is eventually segregated within membrane-enclosed organelles such as vacuoles, multivesicular bodies, and dense bodies, where the molecules may assume a crystalline packing. As a result of the accumulation of ferritin within these inclusions and within filler substance, only a small amount of protein remains to enter the underlying parenchyma. Presentation of ferritin to prefixed cells leads to a random dispersion of free cytoplasmic ferritin. This artifactual distribution in both prefixed and postfixed cells is concurrent with disruption of cell membranes.     

EVIDENCE FROM ELECTRON MICROGRAPHS FOR THE PASSAGE OF MATERIAL THROUGH PORES OF THE NUCLEAR MEMBRANE

1. The nurse cells of Rhodnius possess nucleoli that stain with Heidenhain's hematoxylin but give a negative Feulgen reaction. In localized positions adjacent to the nuclear membrane are seen masses of material both within the nucleus and the adjoining cytoplasm that stain with Heidenhain's hematoxylin, but, like the nucleolus, give a negative Feulgen reaction. 2. Electron micrographs of the nurse cells of Rhodnius reveal the nuclear membrane to contain pores approximately 400 A in diameter. 3. In electron micrographs the nucleolus is seen to be composed of a reticulum containing tightly packed granules. Between the centrally located nucleolus and the nuclear membrane are observed relatively small bunches of granules of the same relative size as those occurring in the nucleolus. Aggregated at certain positions adjacent to the nuclear membrane both within the nucleus and in the adjoining cytoplasm are irregularly shaped masses of granules. Certain of these masses within the nucleus are seen to be continuous with those in the cytoplasm through narrow isthmuses of material extending through pores of the nuclear membrane. Other masses of granules show evidence of preparing to enter the pores by projecting tongues of material toward and into them. In the adjacent cytoplasm pear-shaped masses of granules are seen in front of and in contact with the pores which suggests that they were fixed in the process of or just after completing passage through the pores.     

THE EFFECT OF THE ELECTRON-OPAQUE PORE MATERIAL ON EXCHANGES THROUGH THE NUCLEAR ANNULI

To investigate the extent to which the electron-opaque pore material can regulate nucleocytoplasmic exchanges which occur through the nuclear annuli, experiments were performed in which polyvinylpyrrolidone (PVP)-coated colloidal gold particles (25 to 170 A in diameter) were microinjected into the cytoplasm of amebas (Amoeba proteus). The cells were fixed at various times after injection and examined with the electron microscope in order to determine the location of the gold particles. High concentrations of gold were found associated with the pore material at specific points adjacent to and within the pores. It is tentatively suggested that such specific accumulations could be a means of selecting substances from the cytoplasm for transport through the pores. Particles were also scattered throughout the ground cytoplasm and nucleoplasm. A comparison of the diameters of particles located in these two regions showed that the ability of materials to penetrate the nuclear envelope is a function of their size. It was estimated that the maximum size of the particles able to enter the nucleus is approximately 125 to 145 A indiameter. The regulation of exchanges with regard to particle size is thought to be dependent on the specific organization of the electron-opaque pore material.     

FURTHER OBSERVATIONS ON THE PHENOMENON OF SECONDARY VACUOLATION IN LIVING CELLS

Invagination of the plasma membrane in plant cells forms peripheral or endocytic structures which often contain a complement of membrane-bound vesicles. These structures, or secondary vacuoles, move with the streaming cytoplasm although their velocities are somewhat slower than that for the various organelles within the cytoplasm. They glide over the nucleus or flow from the peripheral cytoplasm onto a transvacuolar strand and continue unabated along the length of a strand. These structures may detach from the plasma membrane as sacs to become positioned in the cytoplasm directly under the tonoplast and project into the primary vacuole. Some endocytic vacuoles may separate from the peripheral cytoplasm and remain free within the primary vacuole; subsequently they can re-associate with the cytoplasm. While the content and function of these vacuoles are yet to be determined, indirect evidence indicates that they are pinocytic in character since the content of an invagination is confined to the sac upon its detachment from the plasma membrane and is subsequently transported throughout the cell by cyclosis.     

THE INCORPORATION AND FATE OF H3-TYROSINE IN THE HAIR CORTEX OF RATS OBSERVED BY RADIOAUTOGRAPHY

The incorporation of H³-tyrosine into the protein of the cells in the cortex of rat hair has been investigated by radioautography. In growing hairs, radioactivity is found in the matrix, the upper bulb, and the whole of the keratogenous zone up to the fully keratinized part of the shaft, 10 and 30 minutes after an injection of labelled tyrosine. This is unequivocal evidence of protein synthesis at these sites. There is a very precise relationship between the end of protein synthesis and the hardening of the cortical cells at the top of the keratogenous zone. The way in which the silver grains of the radioautographs are clustered indicates that at 30 minutes after the injection the isotope is distributed more evenly in the matrix and upper bulb than in the top of the keratogenous zone. Possibly this reflects a difference, at these sites, in the cell components engaged in protein synthesis, or in the proteins being synthesized. The fully keratinized and hardened part of the hair was not radioactive at 10 and 30 minutes after the injection of H³-tyrosine. The rate at which the radioactivity moves into this region shows that the hair of rats grows 0.9 mm/24 hours. Comparison of the degree of radioactivity along the growing hair in the 30-minute, 12-hour, and 36-hour materials shows conclusively that protein accumulates in the cortical cells during their keratinization. An injection of a labelled amino acid does not behave as an ideal pulse dose; consequently, the grain density over the hair cortex at 36 hours is 100 per cent larger than would be expected if an ideal pulse dose situation existed.     

用ESR自旋捕集法研究吸烟烟气处理的脂质体对大鼠粒细胞产生O_2~-的影响

吸烟烟气能引发卵磷脂脂质体的脂质过氧化。若将吸烟烟气处理20s的脂质体作用完整的大鼠粒细胞(RPN),用ESR自旋捕集方法发现,当作用时间在25min内(脂浓度1.0mg/ml)或脂浓度小于15.0mg/ml(而作用时间均为15min)时,这种过氧化的脂质体能增加RPN呼吸爆发产生O_2~-的量。而没有经吸烟烟气处理的新制脂质体,在脂浓度大于0.2mg/ml(作用时间15min)时,都不同程度地抑制RPN产生O_2~-。     

用ESR自旋捕集法研究吸烟烟气处理的脂质体对大鼠粒细胞产生O_2~-的影响

吸烟烟气能引发卵磷脂脂质体的脂质过氧化。若将吸烟烟气处理20s的脂质体作用完整的大鼠粒细胞(RPN),用ESR自旋捕集方法发现,当作用时间在25min内(脂浓度1.0mg/ml)或脂浓度小于15.0mg/ml(而作用时间均为15min)时,这种过氧化的脂质体能增加RPN呼吸爆发产生O_2~-的量。而没有经吸烟烟气处理的新制脂质体,在脂浓度大于0.2mg/ml(作用时间15min)时,都不同程度地抑制RPN产生O_2~-。     

RADIOAUTOGRAPHIC STUDIES OF THE STEM CELL IN THE THYMUS OF THE IRRADIATED RAT

Radioautographic evidence is presented which characterizes the marrow derived stem cell which promotes thymic recovery following irradiation in the rat. These immigrant cells are similar in morphology to blood monocytes and have been called monocytoid, meaning monocyte-like in appearance. The typical cell had abundant pale staining cytoplasm and a nucleus with many invaginations and folds and a fine chromatin structure. There was no prominent nucleolus. The majority of these cells entered the thymus of the irradiated rat via the blood vessels into the septa and made their way through the connective tissue to the outer cortex. Three distinct morphological cell types appeared to be derived from the immigrant cells. These were fibrocyte-like cells which were located within the septa, macrophages located mainly within the medulla and septa, and large blast cells within the cortex, which proliferated giving rise to large thymocytes. The blast cells were characterized as having abundant moderately basophilic (and pyroninophilic) cytoplasm with a distinct cytoplasmic boundary, a large nucleus which still had invaginations and folds, a loose chromatin structure and one or more very prominent nucleoli. They were located in groups primarily within the outer cortex and often adjacent to blood vessels. They were found to be highly susceptible to damage in smear preparations. In contrast, their progeny, the large thymocytes were not highly susceptible to damage in smear preparations but teased out as large round cells with a highly basophilic rim of cytoplasm. The large thymocytes were precursors to medium and small cells. A radioautographic technique for 1 μ tissue sections is also described.     

THE EFFECTS OF VARIATIONS IN THE CONCENTRATION OF OXYGEN AND OF GLUCOSE ON DARK ADAPTATION

In this study we have analyzed the effects of variations in the concentrations of oxygen and of blood sugar on light sensitivity; i.e. dark adaptation. The experiments were carried out in an air-conditioned light-proof chamber where the concentrations of oxygen could be changed by dilution with nitrogen or by inhaling oxygen from a cylinder. The blood sugar was lowered by the injection of insulin and raised by the ingestion of glucose. The dark adaptation curves were plotted from data secured with an apparatus built according to specifications outlined by Hecht and Shlaer. During each experiment, observations were first made in normal air with the subject under basal conditions followed by one, and in most instances two, periods under the desired experimental conditions involving either anoxia or hyper- or hypoglycemia or variations in both the oxygen tension and blood sugar at the same time. 1. Dark adaptation curves were plotted (threshold against time) in normal air and compared with those obtained while inhaling lowered concentrations of oxygen. A decrease in sensitivity was observed with lowered oxygen tensions. Both the rod and cone portions of the curves were influenced in a similar way. These effects were counteracted by inhaling oxygen, the final rod thresholds returning to about the level of the normal base line in air or even below it within 2 to 3 minutes. The impairment was greatest for those with a poorer tolerance for low O₂. Both the inter- and intra-individual variability in thresholds increased significantly at the highest altitude. 2. In a second series of tests control curves were obtained in normal air. Then while each subject remained dark adapted, the concentrations of oxygen were gradually decreased. The regeneration of visual purple was apparently complete during the 40 minutes of dark adaptation, yet in each case the thresholds continued to rise in direct proportion to the degree of anoxia. The inhalation of oxygen from a cylinder quickly counteracted the effects for the thresholds returned to the original control level within 2 to 3 minutes. 3. In experiments where the blood sugar was raised by the ingestion of glucose in normal air, no significant changes in the thresholds were observed except when the blood sugar was rapidly falling toward the end of the glucose tolerance tests. However, when glucose was ingested at the end of an experiment in low oxygen, while the subject remained dark adapted, the effects of the anoxia were largely counteracted within 6 to 8 minutes. 4. The influence of low blood sugar on light sensitivity was then studied by injecting insulin. The thresholds were raised as soon as the effects of the insulin produced a fall in the blood sugar. When the subjects inhaled oxygen the thresholds were lowered. Then when the oxygen was withdrawn so that the subject was breathing normal air, the thresholds rose again within 1 to 2 minutes. Finally, if the blood sugar was raised by ingesting glucose, the average threshold fell to the original control level or even below it. 5. The combined effects of low oxygen and low blood sugar on light sensitivity were studied in one subject (W. F.). These effects appeared to be greater than when a similar degree of anoxia or hypoglycemia was brought about separately. 6. In a series of experiments on ten subjects the dark adaptation curves were obtained both in the basal state and after a normal breakfast. In nine of the ten subjects, the food increased the sensitivity of the subjects to light. 7. The experiments reported above lend support to the hypothesis that both anoxia and hypoglycemia produce their effects on light sensitivity in essentially the same way; namely, by slowing the oxidative processes. Consequently the effects of anoxia may be ameliorated by giving glucose and the effects of hypoglycemia by inhaling oxygen. In our opinion, the changes may be attributed directly to the effects on the nervous tissue of the visual mechanism and the brain rather than on the photochemical processes of the retina.     

Effects of Ethamivan in Patients with Chronic Respiratory Disease

Nineteen patients suffering from chronic respiratory disease were evaluated before, during and after ethamivan administration by serial measurement of arterial pH, pCO₂, plasma ethamivan levels and alveolar ventilation. Ethamivan was administered intravenously as a single injection of 50 mg. in five patients; as an injection of 25 mg./kg. in five patients; as an intravenous injection of (a) 50 mg. over 15 minutes and (b) 150 mg. over 15 minutes in five patients; and finally as an oral dose of 300 to 500 mg. in five patients.Plasma levels of ethamivan became unmeasurable within 15 minutes of receiving the largest dose. Alveolar ventilation increased only in patients receiving the highest intravenous dose, and no significant changes in blood gases were elicited in any patient.     

催产素对豚鼠听觉机能作用的电生理研究

本实验利用听觉电生理学方法,研究了催产素(Oxytocin)对豚鼠内耳听觉机能的作用。给豚鼠肌内注射催产素后,由短声引起的耳蜗微音器电位和听神经复合动作电位幅值增加,听神经复合动作电位和听皮层诱发电位的阈值降低。说明催产素具有提高豚鼠内耳听觉机能的作用。     

THE FINE STRUCTURE OF VON EBNER''S GLAND OF THE RAT

The fine structure of von Ebner's gland was studied in untreated rats and rats stimulated to secrete by fasting-refeeding or injection of pilocarpine. Cytological features were similar to those reported for pancreas and parotid gland. Abundant granular endoplasmic reticulum filled the basal portion of the cell, a well-developed Golgi complex was located in the vicinity of the nucleus, and the apical portion of the cell was filled with dense secretory granules. Dense heterogeneous bodies resembling lysosomes were closely associated with the Golgi complex. Coated vesicles were seen in the Golgi region and also in continuity with the cell membrane. Granule discharge occurred by fusion of the granule membrane with the cell membrane at the secretory surface. Successive fusion of adjacent granules to the previously fused granule formed a connected string of granules in the apical cytoplasm. Myoepithelial cells were present within the basement membrane, and nerve processes were seen adjacent to acinar and myoepithelial cells. Duct cells resembled the intercalated duct cells of the major salivary glands.