Senescence in cell oxidative status in two bird species with contrasting life expectancy

Oxidative stress occurs when the production of reactive oxygen species (ROS) by an organism exceeds its capacity to mitigate the damaging effects of the ROS. Consequently, oxidative stress hypotheses of ageing argue that a decline in fecundity and an increase in the likelihood of death with advancing age reported at the organism level are driven by gradual disruption of the oxidative balance at the cellular level. Here, we measured erythrocyte resistance to oxidative stress in the same individuals over several years in two free-living bird species with contrasting life expectancy, the great tit (known maximum life expectancy is 15.4 years) and the Alpine swift (26 years). In both species, we found evidence for senescence in cell resistance to oxidative stress, with patterns of senescence becoming apparent as subjects get older. In the Alpine swift, there was also evidence for positive selection on cell resistance to oxidative stress, the more resistant subjects being longer lived. The present findings of inter-individual selection and intra-individual deterioration in cell oxidative status at old age in free-living animals support a role for oxidative stress in the ageing of wild animals.     

The postnatal development of the submandibular gland of the mouse

Summary The postnatal development of the submandibular gland was investigated in male mice of the Swiss-Webster strain, which were killed at 1, 2, 3, 4, 5, 6, 8, 10, 12, 16 and 20 weeks of age, while the older mice had been weaned at 3 weeks of age. The mean weight of the submandibular gland increases from 9.5 mg at 1 week to 232.9 mg at 20 weeks of age, and the rate of increase is rapid between 3 and 10 weeks of age. The gland's contents of DNA, RNA and protein increase in a similar manner.The changes in the constituent cell types of the gland were studied in radioautographs prepared from Epon-embedded sections of mice given ³H-thymidine and stained with toluidine blue. At 1 week of age, the gland consists of acinar cells (36%), intercalated duct cells (26%), juxta-acinar cells (13%), striated duct cells (12%) and others. The cellular composition of the gland changes little before weaning, but the absolute number of all types of cells increases with age. Between 3 and 4 weeks, juxta-acinar cells disappear and granular convoluted tubule cells appear and increase rapidly in number with age. The rapid expansion of the population size of granular convoluted tubule cells after weaning coincides with the second peak of increased proliferative activity of intercalated duct cells, whereas all the other cell types show a progressive decrease in their proliferative activity with age. In spite of the burst in proliferative activity, there is no corresponding increase in the absolute number of intercalated duct cells. The number of striated duct cells peak at 5 weeks of age and then declines. These findings indicate that the mitoses of intercalated duct cells give rise to granular convoluted tubule cells through a stage of striated duct cells. At 20 weeks of age, the gland consists of granular convoluted tubule cells (47%), acinar cells (28%), intercalated duct cells (12%), striated duct cells (1%) and others.Supported by Public Health Service Research Grant AMDE 19753 from the National Institute of Health. The authors are indebted to Mr. I. Borcsanyi for technical assistance     

Ultrastructure of the kidney of a South American caecilian,Typhlonectes compressicaudus (Amphibia,Gymnophiona)

Summary The ultrastructure of the distal nephron, the collecting duct and the Wolffian duct was studied in a South American caecilian, Typhlonectes compressicaudus (Amphibia, Gymnophiona) by transmission and scanning electron microscopy (TEM, SEM). The distal tubule (DT) is made up of one type of cell that has a well-developed membrane labyrinth established both by interdigitating processes and by interlocking ramifications. The processes contain large mitochondria, the ramifications do not. The tight junction is shallow and elongated by a meandering course. The connecting tubule (CNT) is composed of CNT cells proper and intercalated cells, both of which are cuboidal in shape. The CNT cells are characterized by many lateral interlocking folds. The intercalated cells have a dark cytoplasm densely filled with mitochondria. Their apical cell membrane is typically amplified by microplicae beneath which a layer of globular particles (studs) is found. The collecting duct (CD) is composed of principal cells and intercalated cells, again both cuboidal in shape. The CD epithelium is characterized by dilated intercellular spaces, which are often filled with lateral microfolds projecting from adjacent principal cells. The apical membrane is covered by a prominent glycocalyx. The intercalated cells in the CD are similar to those in the CNT. The Wolffian duct (WD) has a tall pseudostratified epithelium established by WD cells proper, intercalated cells and basal cells. The WD cells contain irregular-shaped dense granules located beneath the apical cell membrane. The intercalated cells of the WD have a dark cytoplasm with many mitochondria; their nuclei display a dense chromatin pattern.Research fellow of the Alexander von Humboldt Foundation     

青藏高原纳木错高寒草甸生态系统碳交换对多梯度增水的响应

高寒草甸是青藏高原的主要草地类型, 对青藏高原生态系统碳收支具有重要的调节作用。目前, 有关高寒草甸生态系统碳交换对气候变化的响应所知甚少, 尤其是降水变化会如何影响高寒草甸碳交换过程的相关研究非常匮乏。该文作者于2013和2014年的生长季(5-9月)在青藏高原纳木错地区高寒草甸进行多梯度人工增水实验, 设置对照和5个水分添加梯度, 分别增加0%、20%、40%、60%、80%和100%的降水, 以研究高寒草甸生态系统在不同降水量条件下的碳交换变化。增水处理后, 各处理梯度之间的土壤温度没有显著差异, 而土壤含水量在不同增水处理后出现显著变化, 相对于对照, 增水幅度越大, 对应的土壤含水量越高。综合2013和2014年的观测结果, 高寒草甸生态系统整体表现为碳吸收, 在20%增水处理中, 净生态系统碳交换(NEE)达到最大值, 随着模拟的降水梯度进一步增加, NEE逐渐下降; 增水处理对生态系统呼吸(ER)无显著影响; 总生态系统生产力(GEP)的变化趋势与NEE一致, 即随着增水梯度增大, GEP先增加, 并在增水20%处理达到最大值, 随后GEP开始降低。研究表明, 在高寒草甸生态系统, 水分是影响GEP和NEE的重要因素, 对ER影响较弱; 未来适度的增水(20%-40%)能促进高寒草甸生态系统对碳的吸收。     

Responses of ecosystem carbon exchange to multi-level water addition in an alpine meadow in Namtso of Qinghai-Xizang Plateau,China

高寒草甸是青藏高原的主要草地类型, 对青藏高原生态系统碳收支具有重要的调节作用。目前, 有关高寒草甸生态系统碳交换对气候变化的响应所知甚少, 尤其是降水变化会如何影响高寒草甸碳交换过程的相关研究非常匮乏。该文作者于2013和2014年的生长季(5-9月)在青藏高原纳木错地区高寒草甸进行多梯度人工增水实验, 设置对照和5个水分添加梯度, 分别增加0%、20%、40%、60%、80%和100%的降水, 以研究高寒草甸生态系统在不同降水量条件下的碳交换变化。增水处理后, 各处理梯度之间的土壤温度没有显著差异, 而土壤含水量在不同增水处理后出现显著变化, 相对于对照, 增水幅度越大, 对应的土壤含水量越高。综合2013和2014年的观测结果, 高寒草甸生态系统整体表现为碳吸收, 在20%增水处理中, 净生态系统碳交换(NEE)达到最大值, 随着模拟的降水梯度进一步增加, NEE逐渐下降; 增水处理对生态系统呼吸(ER)无显著影响; 总生态系统生产力(GEP)的变化趋势与NEE一致, 即随着增水梯度增大, GEP先增加, 并在增水20%处理达到最大值, 随后GEP开始降低。研究表明, 在高寒草甸生态系统, 水分是影响GEP和NEE的重要因素, 对ER影响较弱; 未来适度的增水(20%-40%)能促进高寒草甸生态系统对碳的吸收。     

Supramol--a program for structure analysis of intercalates using molecular simulations: the structure of VOPO4*C6H4O2

A method of structure analysis of intercalates has been developed that uses a combination of molecular simulations with powder diffraction. The program Supramol for the determination of intercalated structures uses crystal energy minimization in conjunction with powder diffraction data.The program solves the multiple minima problem in molecular mechanics, generating initial models systematically and searching for the global energy minimum by comparing the experimental and calculated diffraction patterns. The program is compatible with the Cerius2 modeling environment.Two intercalated crystal structures solved by Supramol are presented in the present paper: vanadyl phosphate intercalated with p-benzoquinone and the high temperature phase of vanadyl phosphate intercalated with dioxane. The structure of vanadyl phosphate intercalated with p-benzochinone is tetragonal, space group I4/ m, the unit cell parameters a=6.21 A, b=6.21 A, c=20.18 A and the density is rho=2.30 g x cm(-3), Z=4. The crystal structure of vanadyl phosphate intercalated with dioxane (high temperature phase) is monoclinic, space group C2/ m, unit cell parameters are: a= b=8.94 A, c=8.22 A, alpha=gamma=90 degrees, beta=106.30 degrees, Z=4, density 2.248 g x cm(-3).     

Habitat degradation and trout stocking can reinforce the impact of flash floods on headwater specialist Alpine bullhead Cottus poecilopus – A case study from the Carpathian Mountains

In freshwater streams, flooding is a typical source of natural disturbance that plays a key role in the dynamics of animal populations and communities. However, habitat degradation and fish stocking might increase the severity of its impact. We tested the effects of a flash flood on the abundance of three size classes of headwater dwelling Alpine bullhead, Cottus poecilopus, in the streams of the Carpathian Mountains in the Czech Republic, that are stocked with hatchery‐reared brown trout, Salmo trutta. We showed that the overall abundance of Alpine bullhead was highest at the sites with the least degraded habitat (i.e., natural habitat) and we caught almost no Alpine bullhead at the sites with the most degraded habitat. The flash flood had a strong negative effect on the abundance of the largest individuals of Alpine bullhead. Abundance of small and medium size Alpine bullhead was negatively affected by the abundance of adult stocked brown trout before as well as after the flash flood. However, negative effect of adult brown trout abundance on abundance of large Alpine bullhead was not significant before the flash flood, and it became significant after the flash flood. This could indicate an accumulation of negative impacts of trout stocking and flash flood on this size class. Overall, our results suggest that stocking of hatchery trout and habitat degradation can reinforce the impact of flash floods on the population of Alpine bullhead in the streams of the Carpathian Mountains.     

A comparative analysis of the foraging behaviour of the Chough Pyrrhocorax pyrrhocorax and the Alpine Chough Pyrrhocorax graculus coexisting in the Alps

The foraging behaviour of the Chough Pyrrhocorax pyrrhocorax and the Alpine Chough Pyrrhocorax graculus was studied during summer and autumn in the western Italian Alps. We assessed feeding times, feeding rates and foraging techniques associated with different foraging habitats. The Alpine Chough is mainly a ground surface feeder, stays for a rela-tively short time at a feeding site (on average 2.1 min) and feeds quickly (on average 9.2 items/min). In contrast, the Chough is almost exclusively an undersurface feeder (digger and prober), stays at a feeding site twice as long as the Alpine Chough (5.4 min) and feeds four times as slowly (2.2 items/min). These differences suggest that the degree of actual ecological overlap is almost as low as possible for two species using the same Alpine pastures as foraging sites. Interspecific coexistence has occurred through a clear differentiation of foraging strategies and diets. The foraging efficiency (in terms of feeding rate)of the Alpine Chough was always higher than that of the Chough in all habitats where they occurred together. The Alpine Chough was more variable in the use of foraging techniques and more diversified in the use of foraging habitats than the Chough. In both species, juveniles fed less efficiently than adults; the foraging behaviour of the Chough is probably more difficult to learn than that of the Alpine Chough. Other data have also shown that the Alpine Chough is more opportunistic than the Chough in using seasonally available food. Considering the above, we suggest that the foraging behaviour of the Alpine Chough is more flexible and, perhaps, better adapted to the high mountain Alpine environment than that of the Chough. Two hypotheses concerning the ultimate reason why the Alpine Chough and the Chough have evolved divergent beak morphology are discussed.     

Radiation injury of hemopoiesis in alpine conditions depending upon the duration of adaptation

A study was made of the disturbance of haemopoiesis in the small laboratory animals and dogs kept in Alpine conditions (3200 m) and exposed to ionizing radiation on days 3, 15, 22, 25 and 33 of adaptation. The radiation damage to haemopoiesis in Alpine conditions was shown to decrease at the beginning of the adaptation due to the intensification of the regenerating processes, which was manifested by the increase in the rate of DNA synthesis in the haemopoietic organs, and activation of erythropoiesis, myelopoiesis and lymphopoiesis. Later (after 30-35 days), a 2-4-fold increase was noted in the number of haemopoietic stem cells which improved the compensatory potency of the tissue under study and increased the total resistance of the organism.     

A morphometric and ultrastructural study of lithium-induced changes in the medullary collecting ducts of the rat kidney

Summary Rats were given a lithium-containing diet (40 mmol/kg) to Study the effect of lithium on the structure of collecting ducts from the inner stripe of the outer medulla. The results show that there is a significant increase in the volume density of collecting ducts already after one week on this diet. The volume density of both intercalated and principal cells increases, whereas the volume density of mitochondria in the cytoplasm increases in the intercalated cells only. The increased volume of both principal and intercalated cells seems to be part of a general hyperplasia and hyperactivity of the collecting duct, which may in some way be related to the effects of lithium on vasopressinmediated water transport. The specific changes in the intercalated cells may be a consequence of the effects of lithium on distal nephron potassium and hydrogen ion transport in the distal nephron.     

Membrane-associated carbonic anhydrase activity in the kidney of CA II-deficient mice.

Carbonic anhydrase II-deficient mice offer a possibility to study the localization along the nephron of membrane-associated carbonic anhydrase (CA) activity without interference from the cytoplasmic enzyme. We studied the localization of CA in kidneys from CA II-deficient and control mice by immunocytochemistry (CA II) and histochemistry. Cytoplasmic staining was found in convoluted proximal tubule, thick limb of Henle, and principal and intercalated cells of collecting duct in the control animals but was absent in the CA II-deficient mice. In cells with cytoplasmic staining the cell nuclei were stained. Intense histochemical activity was associated with apical and basolateral membranes of convoluted proximal tubule, first part of thin limb, thick limb, and basolateral membranes of late distal tubule. In collecting ducts of control animals, the basolateral cell membranes of intercalated cells were the only clearly stained membranes. In CA II-deficient animals one type of intercalated cell was stained most intensely at the apical membranes and another only at the basolateral. We suggest that the former corresponds to Type A intercalated cells secreting H+ ions to the luminal side and the latter to Type B cells secreting H+ ions to the basolateral side.     

The versatile stellate cell - more than just a space-filler

Most epithelia contain multiple cell types that interact to perform the roles required of the tissue. In insect epithelia, the apical plasma membrane V-ATPase dominates ion-transport models, and (as in vertebrates) is usually found in specialized intercalated cell types or regions. The Malpighian tubules of several insect Orders contain not just a mitochondrion-rich principal cell expressing high levels of V-ATPase, but a smaller, intercalated "type II", "secondary" or "stellate" cell. Recent data show that this cell type plays a key role in control of chloride and water flux across the tissue, but also may play other, still unsuspected dynamic roles.     

AMPK regulates cell shape of cardiomyocytes by modulating turnover of microtubules through CLIP‐170

AMP‐activated protein kinase (AMPK) is a multifunctional kinase that regulates microtubule (MT) dynamic instability through CLIP‐170 phosphorylation; however, its physiological relevance in vivo remains to be elucidated. In this study, we identified an active form of AMPK localized at the intercalated disks in the heart, a specific cell–cell junction present between cardiomyocytes. A contractile inhibitor, MYK‐461, prevented the localization of AMPK at the intercalated disks, and the effect was reversed by the removal of MYK‐461, suggesting that the localization of AMPK is regulated by mechanical stress. Time‐lapse imaging analysis revealed that the inhibition of CLIP‐170 Ser‐311 phosphorylation by AMPK leads to the accumulation of MTs at the intercalated disks. Interestingly, MYK‐461 increased the individual cell area of cardiomyocytes in CLIP‐170 phosphorylation‐dependent manner. Moreover, heart‐specific CLIP‐170 S311A transgenic mice demonstrated elongation of cardiomyocytes along with accumulated MTs, leading to progressive decline in cardiac contraction. In conclusion, these findings suggest that AMPK regulates the cell shape and aspect ratio of cardiomyocytes by modulating the turnover of MTs through homeostatic phosphorylation of CLIP‐170 at the intercalated disks.     

Ultrastructural organization of the transition from the distal nephron to the collecting duct in the desert rodent Psammomys obesus

Summary The transition from the nephron to the collecting duct is formed by three tubular segments (convoluted part of the distal tubule, connecting tubule, cortical collecting duct), which in the desert rodent, Psammomys obesus, transform gradually from one segment to the next, due to intermingling of their different cell types.The convoluted part of the distal tubule (DTC) starts abruptly, shortly beyond the macula densa and initially is homogeneously composed of characteristic DTC-cells. Subsequently, the DTC-cells intermingle with intercalated cells. The first appearance of the connecting-tubule cell, which gradually replaces the DTC-cell, is regarded as the beginning of the connecting tubule. The major portion of the connecting tubule is lined by connecting-tubule cells and intercalated cells. The first appearance of the principal cell between them defines the beginning of the cortical collecting duct, which in the medullary ray is lined by principal and intercalated cells only.Each cell type is described in detail and discussed in relation to the assumed function of the tubular segments.Interspecies differences in the cellular composition of the transitional zone from the nephron to the collecting duct are discussed in relation to the different organization of the collecting duct system.     

Cell type-specific glycoconjugates of collecting duct cells during maturation of the rat kidney

Summary The ontogeny of lectin-positive epithelial cell types and the maturation of polarized expression of the glycocalyx of the collecting ducts (CD) of the rat kidney were studied from samples of 18th-day fetal and neonatal kidneys of various ages. Lectins from Dolichos biflorus (DBA) and Vicia villosa (VVA), with preferential affinity to principal cells, stained virtually all CD cells of the fetal kidneys. However, within two days postnatally, the number of cells positive for DBA and VVA decreased to amounts found in the adult kidneys. Moreover, a characteristic change occurred rapidly after birth in the intracellular polarization of the reactive glycoconjugates, from a uniform plasmalemmal to a preferentially apical staining. In contrast, lectins from Arachis hypogaea (PNA), Maclura pomifera (MPA) and Lotus tetragonolobus (LTA), reacting indiscriminatively with principal and intercalated cells of adult kidneys, stained most CD cells in the fetal kidneys, and failed to show any postnatal change in the amount of positive cells or in the intracellular polarization.The immunocytochemical tests for (Na + K)-ATPase and carbonic anhydrase (CA II) revealed the characteristic postnatal decrease in the amount of principal cells and simultaneous increase in the amount of CA II rich intercalated cells. DBA and VVA reactive cells also decreased postnatally, paralleling the changes observed in the (Na + K)-ATPase positive principal cells. The present results suggest that the expression of the cell type-specific glycocalyx of principal and intercalated cells is developmentally regulated, undergoes profound changes during maturation, and is most likely associated with electrolyte transport phenomena.     

Mountain pasturing of rearing stock reduces the culling risk as dairy cows

Alpine transhumance or droving livestock to mountainous areas during summer months is highly relevant for Austrian agriculture but also for other countries in Alpine regions. Access of rearing stock to mountain pastures is often claimed to be beneficial with respect to health and longevity, but the robust evidence is scarce. Therefore, its effect was tested by including it in the routine genetic evaluation data set for longevity. Alpine transhumance records from 2004 to 2013 were used. After several plausibility checks and restriction to animals with sire and dam known, records of 871 287 dual-purpose Fleckvieh cows sired by 9953 bulls were available. Data were analysed by means of survival analysis accounting for the time-dependent fixed effects of region-year-season, relative performance within herd, change of herd size, and Alpine pasturing of cows, the fixed effects age at first calving and Alpine pasturing of rearing stock, the random time-dependent effect of herd-year and the random genetic effects of sire and maternal grandsire. Fleckvieh cows that had access to Alpine pasture during their rearing period at least once for a minimum of 60 days had functional longevity that was nearly 2 months prolonged compared with cows that had always stayed on the farms as calves or heifers. In a more detailed analysis, the lowest relative culling risk among the significant estimates was observed for cows that had been Alpine pastured in years 1 and 3; it was about 15% below that of cows that never had access to mountain grazing. Evidence for the beneficial effect of Alpine pasturing of rearing stock on the animals’ later fitness, indicated by longevity, could thus be provided.     

Intercalated cells as a probable source for the development of renal oncocytoma

Renal oncocytoma is a distinct type of epithelial tumor said to arise from the collecting duct system. Here we show that in nine of ten oncocytomas the tumor cells expressed an analog of the erythrocyte anion exchanger band 3. In the normal kidney band 3 is confined to the basolateral surface of the majority of intercalated cells which comprise up to 50% of the cortical collecting duct epithelium. Carbonic anhydrase c is another protein abundant in intercalated cells, and this was also expressed in six of the ten oncocytomas investigated. Immunoreactivity specific for band 3 and carbonic anhydrase c was not detected in any of the 20 renal cell carcinomas examined. At favourable section planes direct transitions between normal collecting ducts and oncocytic tubules were observed. These findings suggest that oncocytomas may develop from intercalated cells of the collecting duct epithelium.     

The role of the ultrastructure of the intercalated cells of the kidney collecting tubules in rats in secreting hydrogen ions

Upon experimental metabolic acidosis in rats, induced by a long-term per os administration of a 10% ammonium chloride solution, elevation of titred acidity and ammonium level in rat's urea was registered, in addition to an increased number of intercalated cells in collecting tubules of kidney medullar zone, where the highest carboanhydrase activity occurred. The structural and functional unit of the intercalated cell is a smooth vesicle having presumable H+-,K+-translocating exchange pump within its membrane. The vesicles accumulating gradually H-ions move to the apical cell membrane to release their content by exocytosis into the tubule space. Under H+-ion hypersecretion, upon experimental acidosis, the vesicle membrane is built into the apical plasma membrane, where H+,K+-exchange occurs, thus intensity of this process increasing. The apical plasma membrane of the cell extends its surface at the expense of intracellular canaliculi that are formed.     

Artificial mountain climate in therapy and rehabilitation of patients with chronic nonspecific pulmonary diseases]

Therapeutic efficiency of artificial Alpine climate was studied in a group of 318 patients with chronic nonspecific pulmonary diseases (CNDP). The Alpine climate was modelled on device "Orotron". The base, progradient and prolonged regimes of therapy by artificial Alpine climate (orhotherapy) were used in work. It is shown that application of optimal conditions of orhotherapy promotes normalization of the external respiration function in patients with CNPD due to a decrease in expression of broncho-obstructive and bronchitic syndromes and in nonuniformity of regional ventilation of lungs. Therapy by artificial Alpine climate is recommended to be used in wide clinical practice for treatment and rehabilitation of patients with CNDP.     

Generation of cells with phenotypes of both intercalated duct-type and myoepithelial cells in human parotid gland adenocarcinoma clonal cells grown in athymic nude mice

A neoplastic epithelial cell line was established from nude mice tumors grown after transplantation of surgical specimens from a human parotid gland adenocarcinoma. This cell line, which had ultrastructure similarities to salivary intercalated duct cells, was found by immunohistochemical techniques to contain amylase, but myosin was not detected. Ultrastructurally, cells of an intermediate type between intercalated ductal and myoepithelial cells were found in the transplanted tumors. Moreover, the expression of myosin in addition to the presence of amylase was detected in the tumors. These findings indicate that some transplanted tumor cells appear to be differentiating towards myoepithelial cells.