Interrenal of a female gymnophione amphibian,Ichthyophis beddomei,during the annual reproductive cycle

The interrenal (adrenal) of Ichthyophis beddomei lies on the ventral side of the kidney, distributed in four zones. It is separated from the renal tissue by a thin layer of connective tissue and contains both adrenocortical and chromaffin cells. Adrenocortical tissue constitutes a major portion of the interrenal islets; the chromaffin tissue consists of a few cells located at the peripheries of the interrenal islets. Histochemical studies demonstrate the presence of Δ⁵3β-hydroxysteroid dehydrogenase, 17 β-hydroxysteroid dehydrogenase, glucose-6-phosphate dehydrogenase, succinate dehydrogenase, and sudanophilic lipids in the adrenocortical tissue, suggesting its steroidogenic potential. Annual histometric and histochemical studies show two peaks of interrenal activity: (1) during the breeding phase of the reproductive cycle (January and February) and (2) during the season of heavy monsoon rains (June and July) in the postbreeding phase.     

Functional morphology of thyrotropic and corticotropic cells during reproductive cycle in ewe

A histological examination has been carried on the pars distalis of the pituitaries of twelve, healthy sexually mature balady ewes which were classified into three groups Resting, follicular and luteal phases according to the different phases of reproduction. The histological features of the TSH- ACTH-cells during resting phase has been used to establish a standard with which the subsequent changes during the follicular and luteal phases were compared. Using PAS/OG, AB/OG and PFA/AB/PAS/OG stains the TSH- and ACTH-cells were differentiated and identified from the other forms of the pars distalis cells. The topographical study of the TSH- and ACTH-cells in various regions and levels in the pars distalis of balady ewe as well as their frequencies during the different phase of reproductive cycle were discussed. Statistical analysis of the different cell count during the different phases of reproductive cycle was conducted. TSH- and ACTH-cells exhibited signs of increased cellular activities and significantly increased in number during follicular phase and in the other phases of the reproductive cycle.     

Seasonal study of the adrenal gland of some Indian avian species.

Adrenal glands of eight Indian species of birds, namely Columba livia, Passer domesticus, Corvus splendens, Acridotheres tristis, Acridotheres ginginianus, Milvus migrans, Francolinus pondicerianus and Bubulcus ibis were examined during the sexually active and inactive phases of their annual reproductive cycles. Excepting A ginginianus and M. migrans, among members of either sex of the remaining six species the weight of the adrenal gland increases during the period of sexual activity. Histologically, the interrenal tissue of these birds could be divided into a peripheral subcapsular zone and a central zone. The cytochemical content of these two zones varies between sexual activity and inactivity. In sexually active birds of both sexes, interrenal cells of the central zone exhibit an increased concentration of alkaline phosphatase, glycogen, acid mucopolysaccharides and gross lipids, while in the subcapsular interrenal cells there is a prominent increase of ascorbic acid content. Cytochemical contents of chromaffin cells remain unchanged except acid phosphatase, which increases during the sexually active phase.     

Arylsulphatase A activity and sulphatide concentration in the female rabbit oviduct are under physiological hormonal influence

Summary Oviduct samples of female rabbits in different phases of the reproductive cycle were analysed to establish the role of sex steroid hormones in the regulation of sulphatide concentration and arylsulphatase A activity. In addition to biochemical procedures, histochemical techniques were used to localize both enzyme activity and the natural substrate. The plasma concentrations of progesterone and 17β-oestradiol were determined by radioimmunoassay (RIA). The findings show that the parameters examined undergo considerable changes during the different phases of the reproductive cycle. Oestrogens exert an inducing action on arylsulphatase A activity, while progesterone inhibits it. Fluctuations of the catabolic arylsulphatase activity condition the sulphatide concentration, which reaches maximum values at the eighth post-ovulatory day when progesterone dominance is consolidated. In this phase of the reproductive cycle, thin-layer chromatography confirms the presence not only of larger quantities of sulphatides, but also of all other lipid fractions.     

Melatonin in the regulation of annual testicular events in carp Catla catla: evidence from the studies on the effects of exogenous melatonin, continuous light, and continuous darkness

The physiological significance of melatonin in the regulation of annual testicular events in a major carp Catla catla was evaluated through studies on the effects of graded dose (25, 50, or 100 µg/100 g body wt.) of melatonin exogenously administered for different durations (1, 15, or 30 days) and manipulation of the endogenous melatonin system by exposing the fish to constant darkness (DD) or constant light (LL) for 30 days. An identical experimental schedule was followed during the preparatory (February-March), pre-spawning (April-May), spawning (July-August), and post-spawning (September-October) phases of the annual cycle. Irrespective of the reproductive status of the carp, LL suppressed while DD increased the mid-day and mid-night values of melatonin compared to respective controls. Influences of exogenous melatonin varied in relation to the dose and duration of treatment and the reproductive status of the carp. However, testicular response to exogenous melatonin (at 100 µg, for 30 days) and DD in each reproductive phase was almost identical. Notably, precocious testicular maturation occurred in both DD and melatonin-injected fish during the preparatory phase and in LL carps during the pre-spawning phase. In contrast, testicular functions in both the melatonin-treated and DD fish were inhibited during the pre-spawning and spawning phases, while the testes did not respond to any treatment during the post-spawning phase. In conclusion, this study provided the first experimental evidence that melatonin plays a significant role in the regulation of annual testicular events in a sub-tropical surface-dwelling carp Catla catla, but the influence of this pineal hormone on the seasonal activity of testis varies in relation to the reproductive status of the concerned fish.     

Changes in the Pituitary-interrenal Axis after Gonadectomy in the Catfish,Clarias batrachus (Linn.)

The effects of gonadectomy on the cytological structure of the hypophysis and interrenal tissue of Clarias batrachus have been investigated. Forty days after gonadectomy remarkable hypertrophy of certain cyanophils, identified later as gonadotrops, has been observed followed by hyperplasia. The somatotrops also seem to undergo similar changes, and as a result the proximal pars distalis (PPD) increases enormously and encroaches upon the pars intermedia (PI). The gonadotrops undergo a gradual degranulation and become devoid of stainable cytoplasm by 90 days. No signet ring cells have been noticed at any stage. The cyanophils in the ventral region of the PPD, identified as thyrotrops, also show degranulation, but to a comparatively limited degree. The ACTH cells undergo gradual hypertrophy and degranulation. A progressive increase is noticed in the nuclear diameter. The interrenal cells are activated as indicated by the enhanced nuclear indices. It is suggested that a fall in gonadal steroids may result in ACTH release which in turn activates the interrenal cells.     

Do thyroid and testis modulate the effects of pineal and melatonin on haemopoietic variables inClarias batrachus?

Involvement of pineal and its major hormone, melatonin, in the process of erythropoiesis in a freshwater catfish,Clarias batrachus has been investigated. The study was conducted during four phases, namely preparatory phase, spawning phase, postspawning phase and late postspawning phase of its annual reproductive cycle. During each phase a fish received either melatonin injections or subjected to pinealectomy. In addition, each fish in all the groups, received either iopanoic acid or cyproterone acetate or vehicle in the morning or late afternoon. Results clearly indicate that melatonin stimulates the rate of erythropoiesis inClarias batrachus. It appears that the extent of stimulation depends upon the phase of the annual reproductive cycle. However, in general, the pineal-or melatonin-induced modulation of blood variables is gonad dependent and thyroid seems to play a time of the day dependent subtle role     

Melatonin in the Regulation of Annual Testicular Events in Carp Catla catla: Evidence from the Studies on the Effects of Exogenous Melatonin,Continuous Light,and Continuous Darkness

The physiological significance of melatonin in the regulation of annual testicular events in a major carp Catla catla was evaluated through studies on the effects of graded dose (25, 50, or 100 µg/100 g body wt.) of melatonin exogenously administered for different durations (1, 15, or 30 days) and manipulation of the endogenous melatonin system by exposing the fish to constant darkness (DD) or constant light (LL) for 30 days. An identical experimental schedule was followed during the preparatory (February–March), pre‐spawning (April–May), spawning (July–August), and post‐spawning (September–October) phases of the annual cycle. Irrespective of the reproductive status of the carp, LL suppressed while DD increased the mid‐day and mid‐night values of melatonin compared to respective controls. Influences of exogenous melatonin varied in relation to the dose and duration of treatment and the reproductive status of the carp. However, testicular response to exogenous melatonin (at 100 µg, for 30 days) and DD in each reproductive phase was almost identical. Notably, precocious testicular maturation occurred in both DD and melatonin‐injected fish during the preparatory phase and in LL carps during the pre‐spawning phase. In contrast, testicular functions in both the melatonin‐treated and DD fish were inhibited during the pre–spawning and spawning phases, while the testes did not respond to any treatment during the post‐spawning phase. In conclusion, this study provided the first experimental evidence that melatonin plays a significant role in the regulation of annual testicular events in a sub‐tropical surface‐dwelling carp Catla catla, but the influence of this pineal hormone on the seasonal activity of testis varies in relation to the reproductive status of the concerned fish.     

Phasing in on the cell cycle

Just like all matter, proteins can also switch between gas, liquid and solid phases. Protein phase transition has claimed the spotlight in recent years as a novel way of how cells compartmentalize and regulate biochemical reactions. Moreover, this discovery has provided a new framework for the study of membrane-less organelle biogenesis and protein aggregation in neurodegenerative disorders. We now argue that this framework could be useful in the study of cell cycle regulation and cancer. Based on our work on phase transitions of arginine-rich proteins in neurodegeneration, via combining mass spectroscopy with bioinformatics analyses, we found that also numerous proteins involved in the regulation of the cell cycle can undergo protein phase separation. Indeed, several proteins whose function affects the cell cycle or are associated with cancer, have been recently found to phase separate from the test tube to cells. Investigating the role of this process for cell cycle proteins and understanding its molecular underpinnings will provide pivotal insights into the biology of cell cycle progression and cancer.     

Seasonal changes in the corpuscles of Stannius and the gonads of the catfish, Heteropneustes fossilis (Bloch).

The seasonal changes in the corpuscles of Stannius (CS) and the gonads of the catfish, Heteropneustes fossilis were studied. The annual sex cycle of the fish has been divided into 4 phases on the basis of the variations in the gonosomatic index and histocytological features displayed by the testes and ovaries. There is a rise in the percentage of aldehyde fuchsin (AF)-positive cells in the CS and an increase in the nuclear diameter, at the beginning of preparatory period (February). In the prespawning period (May--June) the AF-positive cells undergo degranulation. A slight regranulation and rise in the percentage of AF-positive cells occurs during early spawing period (July). During the postspawning phase (September--January) the corpuscles remain predominated by AF-negative cells and show histolytical changes; the nuclear indices are reduced. In view of the concomitant changes occurring in the CS and the gonads the possibility of some direct or indirect relationship between the two has been discussed, although it is difficult to ascertain whether the changes in the corpuscles are cause or consequence of the gonadal cycle.     

Effect of isoprenaline on cells in different phases of the mitotic cycle

The effects of isoprenaline on parotid acinar cells in different phases of the mitotic cycle have been investigated. Cells in mitosis at the time of drug administration are not depleted of secretory granules whilst those in other phases are. The drug causes temporary blocks both in metaphase and in the G2 phase. The blocks are prolonged by repeated injections of the drug. Cells continue to undergo DNA synthesis during the period of secretion following the drug. The mitotic delay appears to be specific for the parotid and submaxillary glands.     

Preferential Fas-mediated apoptotic execution at G1 phase: the resistance of mitotic cells to the cell death

Apoptosis is induced by various stresses generated from the extracellular and intracellular environments. The fidelity of the cell cycle is monitored by surveillance mechanisms that arrest its further progression if any crucial process has not been completed or damages are sustained, and then the cells with problems undergo apoptosis. Although the molecular mechanisms involved in the regulation of the cell cycle and that of apoptosis have been elucidated, the links between them are not clear, especially that between cell cycle and death receptor-mediated apoptosis. By using the HeLa.S-Fucci (fluorescent ubiquitination-based cell cycle indicator) cells, we investigated the relationship between the cell cycle progression and apoptotic execution. To monitor apoptotic execution during cell cycle progression, we observed the cells after induction of apoptosis with time-lapse fluorescent microscopy. About 70% of Fas-mediated apoptotic cells were present at G₁ phase and about 20% of cells died immediately after cytokinesis, whereas more than 60% of etoposide-induced apoptotic cells were at S/G₂ phases in random culture of the cells. These results were confirmed by using synchronized culture of the cells. Furthermore, mitotic cells showed the resistance to Fas-mediated apoptosis. In conclusion, these findings suggest that apoptotic execution is dependent on cell cycle phase and Fas-mediated apoptosis preferentially occurs at G₁ phase.     

Diurnal and seasonal profiles of melatonin in the liver and their correlates of ovarian functions under natural photo-thermal conditions in adult carp Catla catla

Present study aimed to demonstrate daily rhythm features of hepatic melatonin concentrations in relation to ovarian functions during four reproductive phases of an annual cycle by measuring the levels of melatonin, 17-β estradiol (E2), vitellogenin (Vg) and maturation inducing hormone (MIH) in the liver and/or serum of adult carp Catla catla. Melatonin titres in liver, irrespective of reproductive phase, underwent daily variations with a peak in early dark phase and nadir at midday. However, the acrophase (Ø) of serum melatonin varied from late night in preparatory phase to midnight in the remaining parts of annual cycle. Their amplitude was highest during post-spawning phase and lowest during spawning phase. Hepatic E2 levels showed daily peak at midday and seasonal peak during pre-spawning phase. Though levels of serum Vg proteins and MIH did not exhibit daily variations, underwent seasonal changes with the highest and lowest values during spawning and post-spawning phases respectively. Hepatic melatonin titres always displayed significant negative correlation with the levels of both E2 and Vg. In essence, our study presented the first data on the daily and seasonal rhythm features of hepatic melatonin in carp and underlined their temporal relationship with the functions of ovary in any fish species.     

Importance of photoperiods in the regulation of ovarian activities in indian major carp Catla catla in an annual cycle

The present study attempted for the first time to explore the importance of photoperiod in the regulation of seasonal ovarian functions in any subtropical major carp. Adult Indian major carp Catla catla were transferred to a long photoperiod (LP; LD 16:8) or a short photoperiod (SP; LD 8:16) for 30 days on 4 dates corresponding to the beginnings of 4 reproductive phases in an annual cycle, and responsiveness of the ovary was evaluated by comparison with the gonadal weight (I(G)), relative number of developing oocytes, serum levels of vitellogenin, and the activity of 2 important steroidogenic enzymes, that is, Delta(5)3beta-hydroxysteroid dehydrogenase and 17.beta-hydroxysteroid dehydrogenase, in the ovary of fish in a natural photoperiod. Exposure of fish to LP during the preparatory phase (February-March) resulted in a significant (p < 0.001) increase in the values of vitellogenin and in the activity of both the steroidogenic enzymes but not in the ovarian weight and in the relative number of different stages of oocytes. A more stimulatory influence of LP was noted during the prespawning phase (April-May), when precocious maturation of ovary was evident from a significant (p < 0.001) rise in the values of each studied features of ovarian functions. However, no ovarian response was found when the fish were transferred to LP during the spawning (July-August) and the postspawning (September-October) phases. On the other hand, the SP was found to have an inhibitory influence on ovarian growth and maturation during the prespawning and the spawning phases or to have no influences on ovarian functions during the preparatory and the postspawning phases of an annual cycle. The results of our study provide the first evidence that photoperiod per se plays an important role in the seasonal maturation of ovary in a subtropical freshwater major carp.     

Effect of sex steroids on the male reproductive organs of the monitor lizard Varanus monitor (Linn.).

Exogenous sex steroids were administered to adult males of the monitor lizard Varanus during the retrogressive and inactive phases of their annual reproductive cycle. Androgen treatment renews spermatogenetic activity and causes an increase in the number of Leydig cells of the testis during the retrogressive phase; during the inactive phase the testicular response to androgen is only slight. In either reproductive phase oestradiol treatment has an inhibitory action on the germ tubules. Progesterone has no effect on the testis in the inactive phase. The vasa deferentia are well developed during the retrogressive phase and thus the effect of androgens is not appreciable. However, during the inactive phase testosterone highly stimulates the deferent ducts. In the inactive phase oestrogen and progesterone also seem to stimulate slightly the deferent tubules; progesterone increases the interstitial tissue of the deferent ducts. Renal sexual segments hypertrophy and become secretory by androgen treatment in either phase of the reproductive cycle, whereas oestrogen and progesterone have no effect. The hemipenes are also stimulated by androgen treatment.     

Cyclic changes in ciliation, secretion and cell height of the oviductal epithelium in women

Oviducts were obtained from women who elected to undergo sterilization either during a normal menstrual cycle, after the first trimester of pregnancy, or in the puerperium. The percent of ciliated cells, cell height and morphology of the fimbria and ampulla were determined and correlated with the stage of the reporductive cycle and plasma levels of the ovarian steroids. Mature ciliated and secretory cells were observed only at mid-cycle. Atrophy, deciliation and loss of secretory activity coincided with elevated levels of serum progesterone. These degenerative processes continued during pregnancy. Ciliation, hypertrophy, and restoration of secretory activity occurred when serum progesterone was essentially undetectable and estradiol relatively low. During each menstrual cycle the secretory cells were observed to undergo a complete cycle of dedifferentiation-differentiation, whereas 10--12% of the ciliated cells lost and regenerated their celia. Ciliogenic cells were frequently present in the epithelium obtained from women in the mid-follicular phase. Fibrous granules, deuterosomes, procentrioles and ciliary buds were observed in the apex of these cells. Plasma levels of estradiol were higher during periods of atrophy and deciliation than they were during periods of hypertrophy and reciliation. It appears that the serum levels of estradiol were adequate to maintain a mature epithelium at all the reproductive stages included in this study. However, progesterone, when present, blocked the growth-promoting effect of estradiol in the oviduct.     

Morphofunctional changes in Leydig cells throughout the continuous spermatogenesis of the freshwater teleost fish, Serrasalmus spilopleura (Characiformes, Characidae): an ultrastructural and enzyme study

The freshwater fish Serrasalmus spilopleura (piranha) has a continuous type of reproduction; gametes are constantly produced and released during the reproductive cycle. The testes do not undergo seasonal morphological changes but exhibit two constant regions throughout the year: the medullar region (involved with spermatogenesis) and the cortical region (involved with spermiation and sperm storage). We have evaluated the ultrastructure of the Leydig cells and the activity of 3beta-HSD (an essential enzyme related to steroid hormone biosynthesis) and acid phosphatase (AcPase; lysosomal marker enzyme) in these two regions. The activity of 3beta-HSD is stronger in the medullar region, and the Leydig cells in this region have a variety of cytological features that reflect differences in hormone synthesis and/or that could be linked to steroidogenic cells under various degrees of hormonal activity. In the cortical region, 3beta-HSD activity is weak and the Leydig cells exhibit signs of degeneration, as confirmed by their ultrastructure and intense AcPase activity. These degenerative signs are indicative of cytoplasmic remodelling to degrade steroidogenic enzymes, such as 3beta-HSD, that could lead to senescence or even to autophagic cell degeneration. S. spilopleura thus constitutes an interesting model for increasing our understanding of steroidogenesis control in freshwater teleost fish.     

The role of miR156 in developmental transitions in Nicotiana tabacum

Plants undergo a series of developmental transitions during their life cycle. After seed germination, plants pass through two distinct phases: the vegetative phase in which leaves are produced and the reproductive phase in which flowering occurs. Based on the reproductive competence and morphological changes, the vegetative phase can be further divided into juvenile and adult phases. Here, we demonstrate that the difference between juvenile and adult phase of Nicotiana tabacum is characterized by the changes in leaf size, leaf shape as well as the number of leaf epidermal hairs(trichomes). We further show that miR156, an age-regulated microR NA, regulates juvenile-to-adult phase transition in N. tabacum. Overexpression of miR156 results in delayed juvenile-to-adult transition and flowering. Together, our results support an evolutionarily conserved role of miR156 in plant developmental transitions.     

Ras is active throughout the cell cycle,but is able to induce cyclin D1 only during G2 phase

The control of cell cycle progression has been studied in asynchronous cultures using image analysis and time lapse techniques. This approach allows determination of the cycle phase and signaling properties of individual cells, and avoids the need for synchronization. In past studies this approach demonstrated that continuous cell cycle progression requires the induction of cyclin D1 levels by Ras, and that this induction takes place during G2 phase. These studies were designed to understand how Ras could induce cyclin D1 levels only during G2 phase. First, in studies with a Ras-specific promoter and cellular migration we find that endogenous Ras is active in all cell cycle phases of actively cycling NIH3T3 cells. This suggests that cyclin D1 induction during G2 phase is not the result of Ras activation specifically during this cell cycle period. To confirm this suggestion oncogenic Ras, which is expected to be active in all cell cycle phases, was microinjected into asynchronous cells. The injected protein induced cyclin D1 levels rapidly, but only in G2 phase cells. We conclude that in the continuously cycling cell the targets of Ras activity are controlled by cell cycle phase, and that this phenomenon is vital to cell cycle progression.     

Histophysiologische Untersuchungen der Entwicklung des Interrenalorgans beim Krallenfrosch (Xenopus laevis Daudin)

Zusammenfassung Mit histologischen und histochemischen Methoden lassen sich bei Xenopus-Larven 4 Zonen des interrenalen Gewebes (Nebennieren-Rinden-Gewebes) in caudo-cranialer Ausdehnung unterscheiden, die verschiedenen Phasen der Nebennierenentwicklung entsprechen. Diese Zonen wurden auf Grund der in ihnen am häufigsten auftretenden Zelldifferenzierungsformen als Blastemzone, Zone der Organbildung, Zone der Ausdifferenzierung und Zone der Funktionsphasen bezeichnet. Während der Entwicklung verändert sich ihre Ausdehnung, sie verschieben sich von cranial nach caudal. Daneben kommt es während der Ausdifferenzierung zu einer Verlagerung der Interrenalzellen von dorsal nach ventral. Die ausdifferenzierten Interrenalzellen durchlaufen einen Funktionszyklus. Charakteristische Differenzierungformen sind hierbei: wenig stimulierte, stark stimulierte, überstimulierte Interrenalzellen sowie Zellen in der Restitutionsform und Degenerationsform.Kerngrößenmessungen und Bestimmungen der Steroiddehydrogenase-Aktivität an Normaltieren zeigen eine Abhängigkeit der Nebennierenaktivität von dem Entwicklungsstadium. Maxima der Aktivität liegen im Stadium 59 und am Ende der Metamorphose vor.Untersuchungen des Organes nach Hypophysektomie und/oder ACTH-Behandlung machen den Einfluß der Hypophyse auf die Normalentwicklung der Nebenniere deutlich. Hypophysektomie verhindert zwar nicht die Abgliederung der Nebenniereninseln vom Blastem, die Zellen differenzieren sich jedoch nicht vollständig aus, sondern verharren in einer funktionellen Ruhepause. ACTH-Behandlung beschleunigt die Differenzierung und funktionelle Entwicklung der Interrenalzellen und führt zu einer Vergrößerung der einzelnen Interrenalkomplexe. Sie kompensiert auch die hemmende Wirkung der Hypophysektomie auf die Nebenniere. Außerdem vermag ACTH die durch Hypophysektomie bewirkte Wachstums-und Entwicklungshemmung teilweise auszugleichen.

---

Histophysiological investigations of the development of the interrenal organ in the clawed toad (Xenopus laevis Daudin)

Summary Histological and histochemical investigations have given evidence for 4 zones of the interrenal tissue in caudo-cranial direction in Xenopus-larvae. These zones correspond to 4 phases of the interrenal development. Due to the representative differentiation form of the interrenal cells they were characterized as: 1) zone of the blastema, 2) zone of the newly formed interrenal organ, 3) zone of differentiation and 4) zone of the various functional stages. During the development these zones shift from cranial to caudal. Besides there is a displacement during the differentiation of the interrenal cells in dorso-ventral direction. Differentiated interrenal cells undergo a functional cycle: hardly stimulated, clearly stimulated and overstimulated adrenocortical cells as well as cells in the state of restitution and degeneration are to be observed.Calculation of the mean nuclear diameter and the activity of the steroiddehydrogenase in normal animals demonstrate a correlation between the activity of the adrenal cortex and the stage of development. Maxima of this activity are found in stage 59 and at the end of metamorphosis. Experiments with hypophysectomy and/or ACTH-treatment clearly demonstrate the influence of the pituitary on the adrenocortex during normal development. Hypophysectomy does not prevent the separation of interrenal islets from the blastema. These cells, however, do not completely differentiate but stay in an inactive phase of their function. ACTH-treatment accelerates the differentiation and functional development of the interrenal cells and further results in an enlargement of the single adrenocortical islets. This treatment compensates the inhibition of the adrenal cortex after hypophysectomy and partly compensates the retardation of development and growth in hypophysectomized animals.

---

---

Mit dankenswerter Unterstützung durch die Deutsche Forschungsgemeinschaft.